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Status |
Public on May 03, 2011 |
Title |
ESC Rep 1 |
Sample type |
RNA |
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Source name |
mES cells Sox1-GFP
|
Organism |
Mus musculus |
Characteristics |
treatment: untreated cell line: 46C mES cells Sox1-GFP differentiation state: Pluripotent
|
Treatment protocol |
ESC: N2B27 media containing LIF and BMP without serum as described previously (Ying and Smith, 2003; Ying et al., 2008). ES cell media was supplemented with the FGF receptor inhibitor PD173074 (Sigma, StemGent) at 100 nM. 48 hr: ES cells were trypsinized, washed, and replated into N2B27 only media for 48 hours.
|
Growth protocol |
Mouse embryonic stem cells were maintained and passaged on gelatin coated plates without feeder cells in N2B27 media containing LIF and BMP without serum as described previously (Ying and Smith, 2003; Ying et al., 2008). ES cell media was supplemented with the FGF receptor inhibitor PD173074 (Sigma, StemGent) at 100 nM to suppress background differentiation. Similar results were obtained using the MEK inhibitor PD0325901 (Sigma, StemGent) at 1 µM. Cells were routinely propagated by trypsinization and re-plating every two to three days, with a split ratio of 1 in 10. N2B27 was prepared with B27 without vitamin A.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted using Trizol, phenol:chloroform extraction, and isopropanol precipitation.
|
Label |
Biotin
|
Label protocol |
Biotinylated and amplified cRNA was prepared by the Illumina TotalPrep RNA Amplification Kit (Applied Biosystems/Ambion).
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|
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Hybridization protocol |
Biotinylated cRNA was hybridized onto Illumina Mouse-Ref8 BeadChip microarrays, according to the manufacturer’s protocol.
|
Scan protocol |
Microarrays were analyzed by an Illumina BeadArray Reader
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Data processing |
raw data were normalized in BeadStudio
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Submission date |
May 02, 2011 |
Last update date |
May 03, 2011 |
Contact name |
Matt Thomson |
E-mail(s) |
thomson@post.harvard.edu
|
Organization name |
Harvard University
|
Department |
MCB
|
Lab |
Ramanathan
|
Street address |
52 Oxford Street 368
|
City |
Cambridge |
State/province |
MA |
ZIP/Postal code |
02138 |
Country |
USA |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE29005 |
Gene expression microarray for mouse ES cells vs. mouse ES cells treated with 48 hr N2B27 |
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