GSM7875029: Peak_3; Mus musculus; ATAC-seq
Approximately 15 to 50 thousand cells were pelleted after FACS isolation by centrifuging at 500g for 5 minutes at 4°C. The pellet was carefully resuspended in 50 μL of cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630), then immediately centrifuged at 4°C, 500g, for 10 minutes. The supernatant was pipetted off, and the pellet was resuspended in 50 uL of transposition buffer (25 uL Tagment DNA Buffer, 2.5 uL Tagment DNA enzyme 1, 22.5 uL nuclease free H2O) and incubated at 37°C for 30 minutes. After transposition, DNA was isolated using the Zymo ChIP DNA Clean & Concentrator Kit per manufacturers instructions, and eluted in 11 uL of 10 mM Tris Buffer, pH 8.0. Library preparation was done using Nextera primer sequences and NEB Next High Fidelity 2X PCR master mix (M0541L) and amplification was run for 10 cycles.