GSM9202190: H146_Anti-H3K4me2_TAS1440_rep1; Homo sapiens; ChIP-Seq
NCI-H1417, NCI-H1417-INSM1KD or NCI-H146 cells were cross-linked at 37°C for 10 min with 1% formaldehyde. After quenching the cross-linking reaction with 1.25 M glycine, cells were lysed in swelling buffer (25 mM HEPES pH 7.9, 1.5 mM MgCl2, 10 mM KCl, and 0.1% (v/v) NP-40) containing a protease inhibitor cocktail (Nacalai Tesque). The mixture was subsequently centrifuged at 5000×g for 5 min at 4°C. Pelleted nuclei were re-suspended in sonication buffer (50 mM HEPES pH 7.9, 140 mM NaCl, 1 mM EDTA, 1% (v/v) Triton X-100, 0.1% (w/v) SDS and 0.1 (w/v) Na-deoxycholate containing a protease inhibitor cocktail, ~5x106 cell/mL), sonicated on a Q700 (QSONICA, Newtown, CT) for 20–25 cycles of 15 s of sonication, followed by 45 s rest, at an amplitude of 60 (total 6000 J). ChIP-seq libraries were constructed using an NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (New England Biolabs, Ipswich, MA, USA) with the SPRIselect reagent (Cat. No.‎ B23318, Beckman Coulter, Indianapolis, IN, USA) according to the manufacturer's protocols.