GSM9085292: K27chipseq WT2, rep2; Neurospora crassa; ChIP-Seq
Fixed cells were lysed using a Branson Sonifier 450 in lysis buffer supplemented with 1 X Halt Protease Inhibitor Cocktail (Thermo Scientific). Chromatin was sheared using a Bioruptor (Diagenode). Immunoprecipitation was performed using anti-H3K27me2/3 antibody (Active motif 39536) or anti-G4 antibody (Sigma-Aldrich MABE917). DNA was purified using Minelute columns (Qiagen). ChIP DNA was subjected to end-repair, adaptor ligation, and PCR amplification using NEBNext Ultra II DNA Library Prep Kit. Libraries were size-selected using AMPure XP beads and sequenced using Illumina NextSeq 500 with either single-end or paired-end reads.