GSM4968824: EP rep2; Mus musculus; ATAC-seq
Cells were collected after sorting into microcentrifuge tubes containing staining media (1xDPBS,1mM EDTA with 5% serum) and centrifuged at 500xg for 5 minutes at 4˚C . Cell pellet was washed with ice-cold 1xDPBS and centrifuged as above. Cells were lysed in (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630) and centrifuged at 500xg for 10 minutes. Samples were suspended 25µL 2xTD Buffer, 2.5µL transposase (Illumina), and 22.5µL nuclease free water and incubated at 37˚C for 30 minutes at 600rpm. Samples were purified using the MinElute Reaction Clean up kit (Qiagen). Library fragments were amplified using NEBnext PCR mastermix and custom Nextera primers, and size selected using Ampure XP Beads.