GSM1314131: H3K36me3 ChIP-seq in KOPT-K1 persisters; Homo sapiens; ChIP-Seq
Chromatin from formaldehyde-fixed cells (1 - 5 x 106 per histone mark, 107 for BRD4) was fragmented to a size range of 200 - 700 bases with a Branson 250 Sonifier. Solubilized chromatin was isolated. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit (Cat #: FC-102-1001). Briefly, DNA was end-repaired using Epicentre’s End-it™ DNA-Repair Kit (Cat #: ER81050). The blunt, phosphorylated ends were treated with Klenow fragment (3 to 5 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of custom adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were isolated using Angencourt® Ampure® XP. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the HiSeq 2500 following the manufacturer's protocols.
GEO Accession
GSM1314131