Illumina HiSeq 2500 paired end sequencing; ATAC-seq of mouse embryonic stem cells
ATAC-seq of mouse embryonic stem cells
mESCs were scraped from the plate and nuclei were isolated using a hypotonic buffer (10 mM Tris-HCl pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% [v/v] Igepal CA-630), and counted. 50,000 nuclei were resuspended in 50 μl of a transposition reaction mix containing 2.5 μl Nextera Tn5 Transposase and 2x TD Nextera reaction buffer. The mix was incubated for 30 min at 37 ºC.  DNA was purified by either the MinElute PCR kit (Qiagen) or the Agencourt AMPure XP (Beckman Coulter) beads and PCR amplified with the NEBNext High Fidelity reaction mix (NEB) to generate DNA libraries.