GSM5820417: P8_1; Mus musculus; ATAC-seq
ATAC-seq was performed as previously described. Briefly, 5¡Á104 cells were harvested for each sample, centrifuged into pellet, and washed in 50¦Ìl cold PBS. Then we gently resuspended the cell pellet in 50¦Ìl cold lysis buffer. The samples were centrifuged immediately at 500 ¡Ág for 10 min, 4¡ãC. After discarded the supernatant, we resuspended the samples in the transposition reaction mix (25 ¦Ìl TD (2x reaction buffer), 2.5 ¦Ìl TDE1 (Nextera Tn5 Transposase), 22.5 ¦Ìl nuclease free H2O), and incubated them at 37¡ãC for 30 min. We purified the transposed DNA with a Qiagen MinElute PCR Purification Kit. The transposed DNA was amplified by PCR. The amplified library was purified with Qiagen MinElute PCR Purification Kit. The quality of purified libraries was further assessed using a Bioanalyzer High Sensitivity DNA Analysis kit (Agilent). Libraries were sequenced on the Illumina HiSeq 2500 with 50 bp paired-end reads and aligned to the reference genome with bowtie2.