GSM4040111: LuxR_ChIP; Vibrio campbellii; ChIP-Seq
Cells were lysed using Bugbuster bacterial lysis reagent coupled with sonication Immunoprecipitated chromatic DNA was subjected to library preparation using NEBNext Ultra II DNA Library kit (NEB cat# E7645S). DNA fragments were end repaired and 5' phosphorylated, and a 3' end Adenosine-tail was added. The products were ligated to a NEB hairpin adapter which contains an uracil at the loop region. The adapter loop was opened by Uracil-Specific Excision Reagent (NEB) to excise the uracil. The products were cleaned with Agencourt AMPure XP beads (Beckman Coulter cat# A63882) but no size selection was applied. 10 (GSF1697) or 12 (GSF2248) PCR cycles were run using 8-nt barcoded oligos from NEBNext Multiplex Oligos barcode kit (NEB cat# 6609S). Libraries were purified with Agencourt AMPure XP beads, multiplexed and sequenced on NextSeq 500 (Illumina) with NextSeq75 High Output v2.5 kit (Illumina cat# 20024906) to generate paired-end 2x42 bp reads.