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Status |
Public on Oct 04, 2012 |
Title |
shCTR biological replicate 4 |
Sample type |
RNA |
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|
Source name |
embryonic stem cell
|
Organism |
Mus musculus |
Characteristics |
tissue: embryonic stem cell transfection: shCTR treatment: none
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNAs were prepared with Qiagen Rneasy minikit according to manufacturer’s protocol
|
Label |
Cy3
|
Label protocol |
500 ng of total RNA from each sample were amplified by Oligo-dT-T7 reverse transcription and labeled by in vitro transcription with T7 RNA polymerase in the presence of Cy3-CTP using the Quick Amp Labeling kit (Agilent) and purified using RNAeasy columns (Qiagen, Hilden, Germany)
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Hybridization protocol |
After fragmentation, 1650 ng of labeled cRNA from each sample was hybridized in in situ hybridization oven (Agilent) for 17 h at 65ºC and washed during 1 min at rt in Gene Expression Wash Buffer 1 (Agilent) and 1 min at 37 ºC with Gene Expression Wahs buffer 2 (Agilent).
|
Scan protocol |
Scanned on an Agilent G2539A scanner at 5um resolution and 100%PMT The intensity data of each individual hybridization were extracted and the quality was assessed with the Feature Extraction software 10.7 (Agilent).
|
Data processing |
Raw data was corrected for background noise using the normexp method. Quantile normalization was applied to assure comparability across samples.
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Submission date |
Oct 02, 2012 |
Last update date |
Oct 04, 2012 |
Contact name |
Luciano Di Croce |
E-mail(s) |
luciano.dicroce@crg.eu
|
Phone |
+34933160132
|
Organization name |
Centre for Genomic Regulation
|
Street address |
Dr. Aiguader, 88
|
City |
Barcelona |
ZIP/Postal code |
08003 |
Country |
Spain |
|
|
Platform ID |
GPL10333 |
Series (1) |
GSE41277 |
Phf19 links methylated lysine 36 of histone H3 to regulation of Polycomb activity |
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