Result

Search results for the GEO ID: GSE17923

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Title

Source name

Description

Characteristics

GSM447605

GPL1261

6114 KSR2-/- Mouse White Adipose Tissue

white adipose tissue, KSR2-/-

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: KSR2-/-

Expression analysis of white adipose tissue from a KSR2-/- mouse.

Sample_geo_accession	GSM447605
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447605/suppl/GSM447605.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

GSM447606

GPL1261

6171 Wild Type Mouse White Adipose Tissue

white adipose tissue, wild type

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: WT

Expression analysis of white adipose tissue from a wild type mouse.

Sample_geo_accession	GSM447606
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447606/suppl/GSM447606.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

GSM447607

GPL1261

6112 KSR2-/- Mouse White Adipose Tissue

white adipose tissue, KSR2-/-

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: KSR2-/-

Expression analysis of white adipose tissue from a KSR2-/- mouse.

Sample_geo_accession	GSM447607
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447607/suppl/GSM447607.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

GSM447609

GPL1261

6169 Wild Type Mouse White Adipose Tissue

white adipose tissue, wild type

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: WT

Expression analysis of white adipose tissue from a wild type mouse.

Sample_geo_accession	GSM447609
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447609/suppl/GSM447609.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

GSM447749

GPL1261

5796 KSR2-/- Mouse White Adipose Tissue

white adipose tissue, KSR2-/-

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: KSR2-/-

Expression analysis of white adipose tissue from a KSR2-/- mouse.

Sample_geo_accession	GSM447749
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447749/suppl/GSM447749.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

GSM447768

GPL1261

5720 Wild Type Mouse White Adipose Tissue

white adipose tissue, wild type

strain: C57Bl6 gender: female age: 6 months tissue: white adipose tissue genotype: WT

Expression analysis of white adipose tissue from a wild type mouse.

Sample_geo_accession	GSM447768
Sample_status	Public on Sep 03 2009
Sample_submission_date	Sep 01 2009
Sample_last_update_date	Sep 02 2009
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	1) Place freshly isolated tissue directly in TriReagent (1ml/50-100mg of tissue).
Sample_extract_protocol_ch1	2) Immediately homogenize tissue and transfer to a sterile 50 ml centrifuge tube.
Sample_extract_protocol_ch1	3) Cap the tube securely, shake, vortex vigorously, and incubate at room temperature 5 min. to assure dissociation of nucleoproteins.
Sample_extract_protocol_ch1	4) Add 0.1 ml BCP/ ml of TriReagent used. Cap the tube, shake, and vortex. Incubate at RT for 10 min. Centrifuge at 3200 rpm in Sorvall tabletop centrifuge or equivalent (1800 g) for 30 min. at 40 C.
Sample_extract_protocol_ch1	5) Of the three phases seen, RNA remains in the topmost aqueous phase. Transfer the aqueous layer to a fresh tube being careful not to disturb the protein-rich interface.
Sample_extract_protocol_ch1	6) Precipitate the RNA by adding isopropanol (molecular biology grade) at 0.5 ml / ml TriReagent used. Shake vigorously, vortex, and leave at room temperature 10-15 min.
Sample_extract_protocol_ch1	7) Centrifuge at 3200 rpm for 45 min. to one hour at 40 C.
Sample_extract_protocol_ch1	8) Decant the isopropanol and wash the pellet gently with 10ml ice cold 75% ethanol (made with 100% ethanol (molecular biology grade) and HPLC grade-H2O). Centrifuge at 3200rpm for 20 min. at 4ºC.
Sample_extract_protocol_ch1	9) Carefully decant the ETOH. Draw off the last with a pipet and allow excess ethanol to evaporate 5-10 min. Do not permit the pellet to completely dry as this will decrease its solubility.
Sample_extract_protocol_ch1
Sample_extract_protocol_ch1	10) Bring the volume of the sample up to 250ul (or appropriate volume) with HPLC-H20. Transfer the RNA sample to a 1.5ml microfuge tube.
Sample_extract_protocol_ch1	11) Add 625μl (2.5X original volume) 100% ice cold ethanol.
Sample_extract_protocol_ch1	12) Add 25μl (0.1X original volume) 3M sodium acetate, pH 5.2.
Sample_extract_protocol_ch1	13) Mix well and place in -200C freezer for 30-60 minutes.
Sample_extract_protocol_ch1	14) Microcentrifuge at top speed for 20-30 min. at 4º C.
Sample_extract_protocol_ch1	15) Decant the ethanol mixture and wash the RNA pellet with 100μl 75% ethanol. Spin at top speed of microfuge for 10 minutes at room temperature. Draw off the supernatant with pipet and repeat the wash step. After drawing off the second 75% wash, invert the tube 5-10 min. to partially dry the sample. Do not completely dry the RNA pellet as this will decrease its solubility.
Sample_extract_protocol_ch1	16) Resuspend in an appropriate amount of HPLC-H2O, depending on the size of the pellet.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	2.5 ug of total RNA was reverse-transcribed and cRNA generated per Affymetrix instructions using the Affymetrix 1-cycle Target Labeling Kit.
Sample_hyb_protocol	Resultant cRNA probes were hybridized to the Affymetrix (Santa Clara, CA) Mouse Genome 430 2.0 Array per manufacturer's suggestion.
Sample_scan_protocol	Following washing and staining, the chips were scanned using a GeneChip Scanner 3,000 6 G in the UNMC microarray core facility
Sample_data_processing	Raw .cel files were processed using Robust Multichip Average (RMA) from GenePattern Software to create a .res file that was subjected to further analysis.
Sample_platform_id	GPL1261
Sample_contact_name	Robert ,,Lewis
Sample_contact_email	kfisher@unmc.edu
Sample_contact_institute	Univ. of Nebraska Medical Center
Sample_contact_address	12706 Lied Transplant Center
Sample_contact_city	Omaha
Sample_contact_state	NE
Sample_contact_zip/postal_code	68105
Sample_contact_country	USA
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM447nnn/GSM447768/suppl/GSM447768.CEL.gz
Sample_series_id	GSE17923
Sample_data_row_count	45101

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