GSM5532176: ICO1-EM-ATAC; Homo sapiens; ATAC-seq
25000 cells were sorted into a 1.5ml tube containing PBS with 5% BSA. 25000 cells were washed once with cold PBS, and then resuspended in Tn5 reaction mix (12.5 µl 2x TD buffer (homemade), 1.25µl Tn5 enzyme (homemade), 0.5 µl Digitonin (Promega, G9441), and 11 µl H2O). Cells were incubated in the thermo mixer at 37 ⁰C, 800 rpm for 20 min. After the incubation, reaction cleanup was performed using QIAGEN MinElute Reaction Cleanup Kit (28204). The purified tagmented DNA samples were barcoded with Nextera Index Kit primers and amplified via PCR followed by a AMpureXP bead (Agencourt, A63882) size selection. Briefly, after first 5 PCR cycles, 5ul of PCR reaction was used for qPCR to determine the number of additional cycles needed. After amplification, PCR reaction was cleaned up with 0.65x AMpureXP beads followed by 1.8x SPRI beads to select DNA fragments from 150 to 600 bp. Finally, libraries were quantified on the bio-analyzer using DNA high-sensitivity kit, and sequenced on an Illumina NextSeq 500 with 38bp paired-end reads at a depth of about 20 million reads per sample.