title: GSM2428963: H2BK5Ac - TSKI4 HDAC6 sh2; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were fixed in 1% paraformaldehyde for 10 minutes, harvested, and the nuclei were isolated and lysed. The nuclear lysate was sonicated and underwent immunoprecipitation using 50 µl of dynabeads (Invitrogen) coupled to 5 µl of Active Motif α-H2BK5Ac antibody per sample. DNA-protein crosslinks were reversed overnight at 65C, and samples were ribonuclease and proteinase treated before DNA was purified using Min Elute columns (Qiagen). Preparation of libraries was performed using 50 ng of ChIP or TSWT input DNA and the KAPA HyperPrep kit with Illumina TruSeq indexed adapters. Dual SPRI size selection was performed after 18 cycles of amplification according to the manufacturer’s protocol. The average library size was ~300 bp. 12-plex libraries were sequenced (1 X 75 bp) using an Illumina NextSeq500 ranging from 7.1-8.9 X 107 reads per sample.
attributes.tag: GEO Accession
attributes.value: GSM2428963