GSM8586833: Hu3112_G0_1_RNAPolIIS5_ChIP_seq; Schizosaccharomyces pombe; ChIP-Seq
After immunoprecipitation, three washing steps were performed using low salt wash buffer (20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 2 mM EDTA (pH 8.0), 1% (w/v) Triton X-100, 0.1% SDS), high salt wash buffer (20 mM Tris-HCl (pH 8.0), 500 mM NaCl, 2 mM EDTA (pH 8.0), 1% (w/v) Triton X-100, 0.1% SDS), and LiCl wash buffer (10 mM Tris-HCl (pH 8.0), 250 mM LiCl, 1 mM EDTA (pH 8.0), 1% IGEPAL CA-630, 0.1% sodium deoxycholate) successively. ChIP-DNA was extracted using the ChIP DNA Clean & Concentrator kit (ZYMO RESEARCH, D5205), and DNA concentration was measured using the Qubit dsDNA HS assay kit (ThermoFisher, Q33231). The sequencing library was prepared using the ThruPLEX DNA-Seq kit (TaKaRa, R400676) with the DNA HT Dual Index Kit – 96N Set A (TaKaRa, R400660). Sequencing was performed using the Illumina Nextseq 2000 platform with the P3 v3 50 kit (36 + 8 + 8 + 36 cycles, single-end sequencing) at the BEA facility (Huddinge, Sweden).