Search results for the GEO ID: GSE9387  |
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|
GSM ID | GPL ID |
Select for analysis |
Title |
Source name |
Description |
Characteristics |
GSM239006 | GPL1355 |
|
Rs256 DMSO 0.001
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239006
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239006/suppl/GSM239006.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239007 | GPL1355 |
|
Rs304 DMSO 0.001
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239007
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239007/suppl/GSM239007.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239008 | GPL1355 |
|
Rs293 DMSO 0.001
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239008
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239008/suppl/GSM239008.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239009 | GPL1355 |
|
Rs293 Myclobutanil 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239009
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239009/suppl/GSM239009.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239010 | GPL1355 |
|
Rs304 Myclobutanil 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239010
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239010/suppl/GSM239010.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239011 | GPL1355 |
|
Rs256 Myclobutanil 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239011
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239011/suppl/GSM239011.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239012 | GPL1355 |
|
Rs256 Myclobutanil 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239012
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239012/suppl/GSM239012.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239013 | GPL1355 |
|
Rs293 Myclobutanil 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239013
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239013/suppl/GSM239013.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239014 | GPL1355 |
|
Rs304 Myclobutanil 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239014
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239014/suppl/GSM239014.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239015 | GPL1355 |
|
Rs293 Myclobutanil 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239015
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239015/suppl/GSM239015.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239016 | GPL1355 |
|
Rs256 Myclobutanil 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239016
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239016/suppl/GSM239016.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239017 | GPL1355 |
|
Rs304 Myclobutanil 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239017
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239017/suppl/GSM239017.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239018 | GPL1355 |
|
Rs256 PB 200uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239018
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239018/suppl/GSM239018.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239019 | GPL1355 |
|
Rs304 PB 200uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239019
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239019/suppl/GSM239019.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239020 | GPL1355 |
|
Rs293 PB 200uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239020
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239020/suppl/GSM239020.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239021 | GPL1355 |
|
Rs293 PCN 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239021
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239021/suppl/GSM239021.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239022 | GPL1355 |
|
Rs304 PCN 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239022
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239022/suppl/GSM239022.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239023 | GPL1355 |
|
Rs293 Propiconazole 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239023
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239023/suppl/GSM239023.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239024 | GPL1355 |
|
Rs256 Propiconazole 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239024
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239024/suppl/GSM239024.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239025 | GPL1355 |
|
Rs304 Propiconazole 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239025
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239025/suppl/GSM239025.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239026 | GPL1355 |
|
Rs256 Propiconazole 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239026
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239026/suppl/GSM239026.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239027 | GPL1355 |
|
Rs293 Propiconazole 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239027
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239027/suppl/GSM239027.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239028 | GPL1355 |
|
Rs304 Propiconazole 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239028
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239028/suppl/GSM239028.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239029 | GPL1355 |
|
Rs304 Propiconazole 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239029
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239029/suppl/GSM239029.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239030 | GPL1355 |
|
Rs293 Propiconazole 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239030
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239030/suppl/GSM239030.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239031 | GPL1355 |
|
Rs256 Propiconazole 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239031
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239031/suppl/GSM239031.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239032 | GPL1355 |
|
Rs256 Triadimefon 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239032
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239032/suppl/GSM239032.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239033 | GPL1355 |
|
Rs304 Triadimefon 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239033
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239033/suppl/GSM239033.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239034 | GPL1355 |
|
Rs293 Triadimefon 10uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239034
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239034/suppl/GSM239034.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239035 | GPL1355 |
|
Rs293 Triadimefon 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239035
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239035/suppl/GSM239035.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239036 | GPL1355 |
|
Rs256 Triadimefon 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239036
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239036/suppl/GSM239036.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239037 | GPL1355 |
|
Rs304 Triadimefon 30uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239037
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239037/suppl/GSM239037.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239038 | GPL1355 |
|
Rs256 Triadimefon 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239038
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239038/suppl/GSM239038.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239039 | GPL1355 |
|
Rs293 Triadimefon 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239039
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239039/suppl/GSM239039.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
GSM239040 | GPL1355 |
|
Rs304 Triadimefon 100uM
|
primary hepatocytes
|
na
|
none
|
| Sample_geo_accession | GSM239040
| | Sample_status | Public on Oct 21 2007
| | Sample_submission_date | Oct 20 2007
| | Sample_last_update_date | Aug 14 2011
| | Sample_type | RNA
| | Sample_channel_count | 1
| | Sample_organism_ch1 | Rattus norvegicus
| | Sample_taxid_ch1 | 10116
| | Sample_treatment_protocol_ch1 | in vitro exposure, 72 hour exposure via medium
| | Sample_molecule_ch1 | total RNA
| | Sample_extract_protocol_ch1 | Total RNA was extracted from primary hepatocytes using TRI Reagent (Molecular Research Center Inc., Cincinnati, OH) according to the manufacturer’s protocol and subjected to quality control measures before application to microarrays. For quality control, RNA A260:A280 ratios were assessed via NanoDrop Fluorospectrometer (NanoDrop Technologies, Inc. Wilmington, DE). RNA absorbance readings with a range 1.8 – 2.1 were followed with DNase treatment, Total RNA Cleanup (Qiagen RNeasy), and then checked for RNA quality using the model 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). Those samples with a ratio of 28S:18S rRNA >1.6 were accepted for subsequent use in DNA microarrays. RNA was stored at -80ºC until labeling for microarray hybridization.
| | Sample_label_ch1 | biotin
| | Sample_label_protocol_ch1 | Five micrograms or purified total RNA from each sample per treatment group was hybridized to Affymetrix GeneChip® Rat Genome 230 2.0 plus microarrays according to the Affymetrix GeneChip Expression Analysis Technical Manual (www.affymetrix.com). Briefly, after purified RNA passed quantity and quality assessment, double stranded cDNA was synthesized from RNA using reverse transcriptase and an oligo-dT primer. Biotinylated RNA (labeled cRNA) served as the microarray target. cRNA was fragmented using heat and magnesium (Mg+2) reducing the cRNA to 25-200 base fragments to facilitate efficient and reproducible hybridization.
| | Sample_hyb_protocol | cRNA was combined with the hybridization cocktail, containing 3nM B2 control oligo, 10 mg/ml Herring Sperm DNA, 50 mg/ml BSA, 100% DMSO, and 2X hybridization buffer (NaCL 5M, MES hydrate Sigma Ultra, MES Sodium Salt, EDTA Disodium Salt 0.5M, 10% Tween-20). GeneChips were hybridized at 45ºC for 16 h. After hybridization the chip was washed and stained with fluorescent streptavidin-phycoerythrin, binding to biotin for detection.
| | Sample_scan_protocol | Signal amplification using anti-Streptavidin antibody (goat) and biotinylated goat IgG antibody was used to bind biotin and provide an amplified fluor that emitted light when the chip is scanned with the GeneChip Scanner 3000.
| | Sample_data_processing | To minimize non-biological factors, e.g. total amount of target hybridized to each array, signal values for each hybridization were multiplied by a scaling factor (SF) to get a trimmed mean intensity equal to 500. Converted .cel files were loaded into JMP Genomics (SAS Inc., Cary, NC), Log2 transformed, normalized using interquartile normalization, and analyzed for significant changes in transcript levels through row-by-row modeling using one-way ANOVA. For initial exploratory analysis, principle component analysis (PCA) was applied using JMP Genomics. Comparisons were made between controls and each treatment group with statistical cut-offs applied at a p-value adjusted false discovery rate and an absolute difference of |1.2| or greater. Probe sets representing transcribed loci, unknown genes, and image clones were removed from the final list of each analysis.
| | Sample_platform_id | GPL1355
| | Sample_contact_name | David,,Dix
| | Sample_contact_email | dix.david@epa.gov
| | Sample_contact_phone | 919-541-2701
| | Sample_contact_department | ORD/NCCT
| | Sample_contact_institute | US EPA
| | Sample_contact_address | 109 TW Alexander Drive
| | Sample_contact_city | Research Triangle Park
| | Sample_contact_state | NC
| | Sample_contact_zip/postal_code | 27711
| | Sample_contact_country | USA
| | Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239040/suppl/GSM239040.CEL.gz
| | Sample_series_id | GSE9387
| | Sample_data_row_count | 31099
| | |
|
| |
| |
Make groups for comparisons |
(2 groups will be compared at a time) |
|
| Select GSMs and click on "Add groups" |
Enter the group name here: |
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