GSM7794004: HeLa EGFP ChIP-Seq; Homo sapiens; ChIP-Seq
The cell lines were crosslinked with 1% formaldehyde for 10 min at room temperature and further processed following the manufacturer's protocol for High Cell number using the kit TruChip Chromatin Shearing Kit, Covaris. Chromatin shearing was performed using Covaris M220 sonicator. 1% of the sheared chromatin was separated for qPCR validation (input sample) and the rest was used for immunoprecipitation. Sonicated chromatin was immunoprecipitated using a goat GFP-antibody (MPI-CBG antibody facility) and Protein G sepharose beads (Protein G Sepharose® 4 Fast Flow, GE Healthcare). Eluates were reverse crosslinked followed by RNA and protein digestion. Sequencing libraries were prepared using NEBNext® Ultra™ DNA Library Prep Kit for Illumina® from 17-75 ng of ChIP DNA with 15 PCR amplification cycle and size selection using AMPure XP beads.