GSM8181424: Rest_D440_Day_0_ATAC; Homo sapiens; ATAC-seq
Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630). Immediately after lysis, nuclei were spun at 500 x g for 10 min in a refrigerated centrifuge. Immediately following the nuclei prep, the pellet was resuspended in the transposase reaction mix (25 µl 2× TD buffer, 2.5 µl transposase (Illumina) and 22.5 µl nuclease-free water). The transposition reaction was carried out for 30 min at 37 °C. Directly following transposition, the sample was purified using a MinElute PCR Purification kit. Library fragments were amplified using 1× NEB next PCR master mix and 1.25 M of custom Nextera PCR primers as previously described, sing the following PCR conditions: 72 °C for 5 min; 98 °C for 30 s; and thermocycling at 98 °C for 10 s, 63 °C for 30 s and 72 °C for 1 min. The libraries were purified using a Qiagen PCR cleanup kit yielding a final library concentration of ∼30 nM in 20 µl.