GSM8329643: TOP1cc-seq WT Input rep1; Homo sapiens; ChIP-Seq
Cell resuspensions were next dounced ten times in a prechilled glass homogenizer. Nuclei were collected and resuspended in shearing buffer (0.1% SDS, 0.5% N-lauroylsarcosine, 1% Triton X-100, 10 mM Tris-HCl pH 8.1, 100 mM NaCl, 1 mM EDTA, 1 mM PMSF, PICs) and incubated on ice for 30 min. Chromatin was fragmented using an E220 Focused-Ultrasonicator (Covaris) to an average size of 200-600 bp. The eluted DNA was quantified with a Qubit 4.0 fluorometer, and 10 ng of DNA was used to prepare the sequencing libraries using KAPA Hyper Prep Kit according to the manufacturer's instructions. Briefly, purified DNA was subjected to end-repair, adaptor ligation followed by indexed PCR. Libraries were size selected for average size of 300 bp. The sequencing was done using Illumina NovaSeq 6000 with a read length configuration of 150 PE for 50 M PE reads per sample (25M in each direction).