title: GSM8439115: MCF7, H3K27me3, in-house pG-Tn5, replicate 1; Homo sapiens; OTHER
library_construction_protocol: MCF7 cells were briefly centrifuged followed by incubation with Nuclear Extraction buffer on ice for 10 minutes. After resuspension in Wash Buffer, nuclei were slow frozen in cryogenic vials with 10% DMSO for storage at –80 °C before resuming the rest of the protocol. Approximately 100,000 cells were used per sample for overnight incubation with primary antibody (1:100 CST #9733T or 1:100 ab4729). Secondary antibody ( 1:100 ABIN101961) was incubated for 30 minutes at room temperature followed by addition of commercial pAG-Tn5 (EpiCypher SKU: 15-1017) or in-house pG-Tn5 preloaded with adapters. Phenol chloroform DNA extraction was performed using Qiagen's Maxtract High Density phase-lock tubes. Libraries were PCR amplified for 12 cycles with Illumina i5 or i7 barcoded primers and Ampure XP beads were used for post-PCR cleanup.