title: GSM6675535: MCF10A_STAG1_CTCF-KD2_ChIP-seq; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were fixed for 15 min in 1% formaldehyde and after quenchig and lysis, sonication with performed in Covaris instrument. Around 50 micrograms of antibody was added to immunoprecipitate chromatin from 10 million cells. 5% mouse ES cell chromatin was added for calibration. At least 5 ng of DNA were processed through subsequent enzymatic treatments with "NEBNext Ultra II FS DNA Library Prep Kit for Illumina" from New England BioLabs (ref. E7805). Briefly, a short fragmentation of 10 min was followed by end-repair, dA-tailing, and ligation to adapters. Adapter-ligated libraries were completed by limited-cycle PCR (8-12 cycles). Resulting average fragment size is 300 bp from which 120 bp correspond to adaptor sequences. Libraries were applied to an Illumina flow cell for cluster generation and sequenced on Illumina NextSeq 500 (with v2.5 reagent kits) following manufacturer's recommendations.