GSM3703336: D31; Homo sapiens; ChIP-Seq
Nucleosomal fraction of the chromatin was digested with 30 U micrococcal nuclease (MNase) for 5 min before incubation overnight with 2µg of antibody and cleaning of the immunoprecipitated chromatin. Libraries were prepared using IP-Star® Compact Automated System (Diagenode Cat# B03000002) from input and ChIP'd DNA (starting amount 1ng) using MicroPlex Library Preparation Kit v2 (12 indices) (Diagenode Cat# C05010013). Library amplification is assessed using High Sensitivity NGS Fragment Analysis Kit (DNF-474) on a Fragment Analyzer™ (Advanced Analytical Technologies, Inc.). Libraries were then double size-selected and purified using Agencourt® AMPure® XP (Beckman Coulter) and quantified using Qubit™ dsDNA HS Assay Kit (Thermo Fisher Scientific, Q32854). Finally their fragment size was analyzed by High Sensitivity DNA Analysis Kit on a 2100 Bioanalyzer system (Agilent).