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Status |
Public on Oct 25, 2012 |
Title |
A3-1 mRNA microarray - siGFP rep2 |
Sample type |
RNA |
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Source name |
cultured mouse embryonic stem cell
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Organism |
Mus musculus |
Characteristics |
sirna sequence: UGAAUUAGAUGGCGAUGUUdTdT/GACAUCGCCAUCUAAUUCAdTdT cell line: A3-1
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Treatment protocol |
Mouse embryonic stem cell were treated with siRNA for GFP or for Lin28a and incubated for 48 hours.
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Growth protocol |
Mouse embryonic stem cell were incubated in DMEM media containing 20% FBS, non-essentail amino acid and LIF at 37˚C.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was purified with Trizol reagent (Invitrogen) according to manufacturer's protocol
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Label |
biotin
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Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 300 ng total RNA (Wt sensetarget label manual, 2007, Affymetrix).
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Hybridization protocol |
Hybridization cocktails containing 5 to 5.5 µg of fragmented, end-labeled single-stranded cDNA were prepared and hybridized to GeneChip Human gene 1.0 ST arrays for 17 hr at 45˚C. GeneChips were washed and stained in the Affymetrix Fluidics Station 450
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Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 7G
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Description |
A3-1 treated with control siRNA
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Data processing |
All gene array data was analyzed using tools in Expression Console software (Affymetrix). The Robust Multi-array Analysis (RMA) (Irizarry, Hobbs, Collin, Beazer-Barclay, Antonellis, Scherf, and Speed 2003) algorithm was used for probeset (gene-level) intensity analysis.
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Submission date |
Apr 09, 2012 |
Last update date |
Oct 25, 2012 |
Contact name |
Hyeshik Chang |
E-mail(s) |
[email protected]
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Organization name |
Seoul National University
|
Department |
School of Biological Sciences
|
Lab |
Hyeshik Chang Lab
|
Street address |
Building 203 Room 525, School of Biological Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu
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City |
Seoul |
State/province |
South Korea |
ZIP/Postal code |
08826 |
Country |
South Korea |
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Platform ID |
GPL6246 |
Series (2) |
GSE37113 |
Transcriptome profiling for changes upon Lin28a knockdown in mouse embryonic stem cell (A3-1) [Affymetrix] |
GSE37114 |
LIN28A targets, regulation of microRNA biogenesis, and effect on transcriptome in A3-1 cells |
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