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Search results for the GEO ID: GSE15581

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GSM389821

GPL1261

Mouse primary megakaryocytes, wild type

Primary megakaryocytes cultured from wild type fetal liver at E14.5

genotype: wild type strain: mixed background gender: female and male age: E14.5 tissue: primary megakaryocytes

Gene expression in primary megakaryocytes cultured from wild type fetal livers.

Sample_geo_accession	GSM389821
Sample_status	Public on Mar 22 2010
Sample_submission_date	Apr 07 2009
Sample_last_update_date	Mar 22 2010
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_biomaterial_provider_ch1	Hozumi Motohashi, Tohoku University Graduate School of Medicine
Sample_treatment_protocol_ch1	None.
Sample_growth_protocol_ch1	Whole livers were recovered from mouse fetuses at E14.5 and single cell suspensions were prepared by successive passage through 25-gauge needles. Fetal liver cells were maintained in RPMI1640 (Wako) supplemented with 20% charcoal-stripped fetal bovine serum, 50 units/ml penicillin, 50 µg/ml streptomycin, and 50 ng/ml of recombinant human thrombopoietin (TPO; generously provided by Kirin). CD41+ cells were selected from a 4 day primary culture of E14.5 fetal liver cells using a MACS magnetic system (Miltenyi Biotec). The fetal liver culture with TPO described above was incubated with FITC-conjugated anti-CD41 antibody (BD Pharmingen, clone MWReg30) followed by incubation with anti-FITC microbeads. Subsequently, the microbeads were selected magnetically through MACS large cell columns (Miltenyi Biotec).
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA was extracted from the CD41+ cells using Isogen (Nippon Gene). RNA was further purified using an RNeasy Mini Kit (Qiagen).
Sample_label_ch1	biotin
Sample_label_protocol_ch1	1µg of total RNA was reverse transcribed using the SuperScript II system for cDNA synthesis (Invitrogen) according to the protocol recommended by Affymetrix. The oligonucleotide used for priming was 5’-ggccagtgaattgtaatacgactcactatagggaggcgg-(t)24-3’ (Affymetrix). Double-stranded cDNA was cleaned up by using the GeneChip® Sample Cleanup Module (Affymetrix). In vitro transcription was performed on all of cDNA using the GeneChip® IVT Labeling Kit (Affymetrix). The cRNA was cleaned up by using RNeasy Mini spin column (QIAGEN). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.
Sample_hyb_protocol	10 µg of fragmented cRNA were hybridized (45°C, 16 hours). Hybridization was controled by adding cRNA cocktail (BioC, BioD, Cre) mixed as described in previous versions of the Expression Analysis Technical Manual. Washing and staining was performed in a Fluidics Station 450 (Affymetrix) using the protocol EukGE-WS2v5.
Sample_scan_protocol	Scanning was performed in an Affymetrix GeneChip® Scanner 3000. Chip analysis was performed using the Affymetrix GeneChip® Operating Software v1.3.
Sample_data_processing	Signal intensity was calculated by MAS5-method (target intensity=100).
Sample_platform_id	GPL1261
Sample_contact_name	Hozumi,,Motohashi
Sample_contact_email	hozumim@med.tohoku.ac.jp
Sample_contact_phone	81-22-717-8087
Sample_contact_fax	81-22-717-8090
Sample_contact_department	Center for Radioisotope Science
Sample_contact_institute	Tohoku University Graduate School of Medicine
Sample_contact_address	2-1 Seiryo-machi, Aoba-ku
Sample_contact_city	Sendai
Sample_contact_state	Miyagi
Sample_contact_zip/postal_code	980-8575
Sample_contact_country	Japan
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM389nnn/GSM389821/suppl/GSM389821.CEL.gz
Sample_series_id	GSE15581
Sample_data_row_count	45101

GSM389826

GPL1261

Mouse primary megakaryocytes, NF-E2p45-deficient

Primary megakaryocytes cultured from NF-E2p45-null fetal liver at E14.5

genotype: NF-E2p45-deficient strain: mixed background gender: female and male age: E14.5 tissue: primary megakaryocytes

Gene expression in primary megakaryocytes cultured from NF-E2p45-null fetal livers.

Sample_geo_accession	GSM389826
Sample_status	Public on Mar 22 2010
Sample_submission_date	Apr 07 2009
Sample_last_update_date	Mar 22 2010
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Mus musculus
Sample_taxid_ch1	10090
Sample_biomaterial_provider_ch1	Hozumi Motohashi, Tohoku University Graduate School of Medicine
Sample_treatment_protocol_ch1	None.
Sample_growth_protocol_ch1	Whole livers were recovered from mouse fetuses at E14.5 and single cell suspensions were prepared by successive passage through 25-gauge needles. Fetal liver cells were maintained in RPMI1640 (Wako) supplemented with 20% charcoal-stripped fetal bovine serum, 50 units/ml penicillin, 50 µg/ml streptomycin, and 50 ng/ml of recombinant human thrombopoietin (TPO; generously provided by Kirin). CD41+ cells were selected from a 4 day primary culture of E14.5 fetal liver cells using a MACS magnetic system (Miltenyi Biotec). The fetal liver culture with TPO described above was incubated with FITC-conjugated anti-CD41 antibody (BD Pharmingen, clone MWReg30) followed by incubation with anti-FITC microbeads. Subsequently, the microbeads were selected magnetically through MACS large cell columns (Miltenyi Biotec).
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA was extracted from the CD41+ cells using Isogen (Nippon Gene). RNA was further purified using an RNeasy Mini Kit (Qiagen).
Sample_label_ch1	biotin
Sample_label_protocol_ch1	1µg of total RNA was reverse transcribed using the SuperScript II system for cDNA synthesis (Invitrogen) according to the protocol recommended by Affymetrix. The oligonucleotide used for priming was 5’-ggccagtgaattgtaatacgactcactatagggaggcgg-(t)24-3’ (Affymetrix). Double-stranded cDNA was cleaned up by using the GeneChip® Sample Cleanup Module (Affymetrix). In vitro transcription was performed on all of cDNA using the GeneChip® IVT Labeling Kit (Affymetrix). The cRNA was cleaned up by using RNeasy Mini spin column (QIAGEN). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.
Sample_hyb_protocol	10 µg of fragmented cRNA were hybridized (45°C, 16 hours). Hybridization was controled by adding cRNA cocktail (BioC, BioD, Cre) mixed as described in previous versions of the Expression Analysis Technical Manual. Washing and staining was performed in a Fluidics Station 450 (Affymetrix) using the protocol EukGE-WS2v5.
Sample_scan_protocol	Scanning was performed in an Affymetrix GeneChip® Scanner 3000. Chip analysis was performed using the Affymetrix GeneChip® Operating Software v1.3.
Sample_data_processing	Signal intensity was calculated by MAS5-method (target intensity=100).
Sample_platform_id	GPL1261
Sample_contact_name	Hozumi,,Motohashi
Sample_contact_email	hozumim@med.tohoku.ac.jp
Sample_contact_phone	81-22-717-8087
Sample_contact_fax	81-22-717-8090
Sample_contact_department	Center for Radioisotope Science
Sample_contact_institute	Tohoku University Graduate School of Medicine
Sample_contact_address	2-1 Seiryo-machi, Aoba-ku
Sample_contact_city	Sendai
Sample_contact_state	Miyagi
Sample_contact_zip/postal_code	980-8575
Sample_contact_country	Japan
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM389nnn/GSM389826/suppl/GSM389826.CEL.gz
Sample_series_id	GSE15581
Sample_data_row_count	45101

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