GSM2474024: PrEC NOMe-seq; Homo sapiens; OTHER
Exponentially growing cells were washed with PBS, trypsinized, and incubated with ice-cold lysis buffer (10 mM Tris, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1 mM EDTA, and 0.5% NP-40) for 5 min on ice to isolate intact nuclei. Nuclei were washed with ice-cold wash buffer (10 mM Tris, pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1 mM EDTA), resuspended in ice-cold 1x GpC buffer (New England BioLabs), and treated with 200 units of M.CviPI enzyme supplemented with1.5 xL S-adenosylmethionine (SAM) for 7.5 min with a boost of 100 units enzyme and 0.75 _L SAM for an additional 7.5 min. Genomic DNA was isolated by standard phenol-chloroform extraction and ethanol precipitation. WGBS libraries were generated using 2-5 µg of DNA and sequenced on the HiSeq2000 performed by the USC Epigenome Center (Blatter et al., Genome Biol. 2014;15:469) WGBS libraries were prepared based on previously described (Berman et al 2012) and according to Illumina's recommendation.
GEO Accession
GSM2474024