GSM8717906: RmpA ChIP sample; Klebsiella pneumoniae; ChIP-Seq
Cells were crosslinked with formaldehyde (1% final concentration) for 20 min at room temperature with continuous shaking. Crosslinking was stopped by addition of 500 mM glycine.The DNA was sonicated to sizes of 200-500 bp on ice (50% total output, 30-s on, 5 min off, twice), following by centrifugation to remove the insoluble cellular debris.The 10% lysis sonicated chromatin was stored and named “Input”, and 80% was used in immunoprecipitation reactions with monoclonal ANTI-FLAG® M2 antibody (Sigma-Aldrich,F1804-50UG) and named “IP”. The high-throughput DNA sequencing libraries were prepared by using VAHTS Universal DNA Library Prep Kit for Illumina V3（Catalog NO. ND607, Vazyme）. The library products corresponding to 200-500 bps were enriched, quantified and finally sequenced on DNBSEQ-T7 sequencer (MGI Tech Co., Ltd. China) with PE150 model.