Cells were transfected with RNaseH1-GFP construct for 48 hrs
source
Fixed chromatin
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
After reversing the cross-links and Proteinase K treatment, the chromatin was purified using the MinElute PCR Purification kit (Cat. 28004, Qiagen) and DNA concentration for library preparation was determined using Qubit fluorometric quantitation (ThermoFisher Scientific). Libraries were prepared from 8ng of DNA using the NEBNext Ultra II DNA library kit for Illumina (Cat. E7645S, NEB), following the manufacturers' instructions. Size selection was performed prior to PCR amplification using RNA clean XP beads (Cat. A63987, Beckman Coulter). Adaptors, PCR amplification and Index Primers were used to multiplex libraries (Multiplex oligos for Illlumina, Cat. E7335S and E7500S, NEB). Libraries were purified using RNA clean XP beads (Cat. A63987, Beckman Coulter) and library size was assessed before high-throughput sequencing by Bioanalyzer (Agilent) using the High Sensitivity DNA analysis kit (Cat. 5067-4626, Agilent). ChIP-seq libraries were sequenced paired-end on an Illumina HiSeq2500 sequencer at the Wellcome Trust Sanger Institute (Cambridge, UK).