title: GSM8878437: ChIP control with untagged cells, Late-G1, Rep1 (Input); Homo sapiens; ChIP-Seq
library_construction_protocol: 20 million cells were collected by trypsinization and resuspended in 1 ml PBS. Cells were crosslinked at room temperature for 10 min by adding an equal volume of 2% PFA. Crosslinked cells were washed with PBS and 0.5% NP-40 in PBS and then lysed in 1 ml ChIP lysis buffer. Chromatin was fragmented using Covaris S2. Lysate was centrifuged at top speed and then pre-cleared with empty protein G dynabeads for at least 1 hour. Cleared lysate was then incubated with FLAG antibody conjugated protein G dynabeads overnight. Beads were washed with a series of washing buffers and then proteins were eluted using 1% SDS in TE buffer. Eluates were reverse crosslinked with proteinase K at 65 ºC for at least 4 hours. DNA was purified using the NucleoSpin Gel and PCR Clean-up Kit. NGS libraries were prepared from 5-10 ng DNA using the NEBNext Ultra II DNA Library Prep Kit for Illumina following the manufacturer's instructions.
identifiers.namespace: GEO
identifiers.id: GSM8878437_r1