GSM8833152: DMSO, input, R1; Homo sapiens; ChIP-Seq
Nuclei were isolated and sonicated using a Covaris ultrasonicator (M220). Each sample was sheared for 30 minutes at following conditions: Duty Factor = 10%, Peak Power = 75.0, Cycles/Burst = 200. Sheared chromatin was incubated with 5µg anti-HA (rabbit CST) per sample overnight at 4°C on a rotator, followed by immunoprecipitation using magnetic beads. After washing, DNA was purified using the Qiagen PCR purification kit (#28104). DNA fragments obtained after ChIP were quantified using Qubit dsDNA HS Assay Kit (Invitrogen). 5ng of DNA was used for library preparation using NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645S), SPRIselect beads (Beckman Coulter, #B23317) and NEBNext Multiplex Oligos for Illumina (NEB, Set 1 #E7335S) following NEB's guidelines (NEB, E7645S). All the libraries were done without size selection with an input of 5ng. ChIP libraries were sequenced as 50bp Paired-End reads on NovaSeq 6K SP.