title: GSM8553191: H1299, DMSO YEATS2 ChIP-seq, rep2; Homo sapiens; ChIP-Seq
library_name: GSM8553191
library_construction_protocol: In ChIP-Seq,H1299 cells that stably expressed N-terminal Flag-YEATS2 were cultured in a 150 mm dish before treating with DMSO or 20 μM inhibitor for 4 hours. Cells were cross-linked by adding formaldehyde (37% v/v) to a final concentration of 1% (v/v) for 10 min at room temperature. After quenching and cell lysis, chromatin was sonicated to 150~500 bp fragment using sonicator. And 100 μg chromatin was diluted to 500 μL using sonication buffer and incubated with 10 μg antibody (anti-FLAG M2 MS) overnight at 4 °C. In RNA-Seq, Total RNA was isolated by Total RNA Isolation Mini Kit (Vazyme). The size distribution and quantity of the DNA library were detected by a bioanalyzer and Qubit. In ChIP-Seq,DNA was purified and then ligated with Illumina adapters. The final DNA library was obtained by size selection and PCR amplification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. In RNA-Seq, the RNA library was prepared by end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.The quantified library was then pooled and sequenced on the Illumina platform NovaSeq PE150.
identifiers.namespace: GEO
identifiers.id: GSM8553191_r1