title: GSM1265859: H3K27me3 ChIPSeq BCBL-1; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were crosslinked with 1% formaldehyde at room temperature for 10 min, and glycine added to 0.125M to terminate crosslinking. Cells were washed twice in ice-cold PBS with protease inhibitors and harvested by centrifugation. Every 2x10E7 cells were lysed in 1 ml ice-cold Farnham lysis buffer (5mM PIPES [pH8.0], 85 mM KCl, 0.5% NP-40) with protease inhibitors. The nuclei were spun down and resuspended in 1 ml RIPA buffer (1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS in 1xPBS) with protease inhibitors. ChIP-enriched DNA was blunt ended with T4 DNA polymerase and Klenow, and phosphorylated with T4 PNK. A 3’ A residue was added with Klenow exo-.  Following adapter ligation, 150 bp to 300 bp DNA fragments were gel-purified, PCR amplified for 18 cycles, quantified with QuantIT dsDNA Assay Kit, and sequenced.
attributes.tag: GEO Accession
attributes.value: GSM1265859