GSM4932377: HEK293T_H3S28ph_dMSK1_non-targeting_gRNAs_re4; Homo sapiens; ChIP-Seq
DNA was fragmented with a bioruptor and chromatin was isolated with indicated antibody. ChIP libraries were prepared using the KAPA Hyper Prep Kit (Roche) following the manufacturer's protocol.  Briefly, 3 ng of fragmented DNA were used to repair the ends before ligation to a diluted NEXTflex DNA adapter (PerkinElmer).  The adaptor-ligated DNA was then enriched using 10 cycles of PCR (1 cycle at 98°C for 45 sec; 10 cycles at 98°C for 15 sec, 60°C for 30 sec, 72°C for 30 sec; and 1 cycle at 72°C for 1 min).  DNA was purified using AMPure XP beads (Beckman Coulter) after adaptor ligation and PCR enrichment, and the libraries were run on a 2200 TapeStation (Agilent Technologies) for quality control.  A KAPA Library Quantification Kit (KAPA Biosystems) was utilized to quantify the libraries for pooling, and a final concentration of 1.5 nM was loaded onto an Illumina cBOT for cluster generation before sequencing on an Illumina HiSeq3000 for sequencing using a Single Read 50 bp run.