Sex: M genotype: WT age: P7 ChIP: 1 chip position: F
Extracted molecule
total RNA
Extraction protocol
TRIzol with O/N precipitation in 100% isopropanol
Label
Cy3
Label protocol
Labeling of high quality samples was performed according to manufacturer’s instructions for the Illumina Total Prep RNA Amplification Kit (Life Technologies, Grand Island, NY). Briefly, 200 ng of total RNA was reverse transcribed with T7 Oligo d(T) to create second strand cDNA. Subsequently, the products were column purified and then in vitro transcribed to generate biotin-labeled cRNA.
Hybridization protocol
cRNA products were column purified and hybridized, per Illuminas protocol, onto Illumina Mouse Ref8 Beadchips for 16 hours at 58° C. Following hybridization, the arrays were washed, stained with streptavidin-cy3 conjugate
Scan protocol
scanned in an Illumina BeadArray Reader
Data processing
The data were normalized using fastlo normalization in R
