GSM7635070: WT_ChIP, rep 3, ChIPseq; Mus musculus; ChIP-Seq
Cultured NK cells from Reg-1fl/fl and Reg-1ΔNK were cross-linked with formaldehyde (final concentration; 1%) followed by quenching with glycine. Wash the cells with PBS and resuspend the cell pellet with cell lysis buffer for ChIP and incubate for 10 min on ice. Pellet the cell nuclei by centrifugation, then resuspend the nuclear pellet in nuclei lysis buffer for ChIP followed by chromatin fragmentation with Bioruptor (Sonicbio Co., Ltd.). Purified ChIP-DNA was applied for library preparation by DNA SMART ChIP-Seq Kit (TaKaRa) following the manufacturer's instructions. Briefly, each 1 ng of purified ChIP-DNA demonstrated denaturation, dephosphorylation of 3'-Ends, and T-Tailing, then dsDNA was produced by Poly(dA) primer annealing and template-switching reaction. The obtained dsDNA was directly amplified into ChIP-seq libraries using SeqAmp DNA Polymerase and the forward and reverse primers from the Indexing Primer Set HT for Illumina followed by size selection using AMPure XP beads (Beckman Coulter), and 100-bp paired-end sequenced using a NovaSeq 6000 (Illumina).