GSM4648814: hMGL SORL1-KO Rep2; Homo sapiens; ATAC-seq
ATAC-seq analysis was performed using the Omni-ATAC-seq protocol (Corces et al., 2017). Briefly, 80,000 viable hMGLs per line were sorted using BD FACSAriaIIu 16-color high-speed cell sorter. Cells were lysed in 50 ml ATAC-Resuspension Buffer containing 0.1% NP40, 0.1% Tween-20, and 0.01% Digitonin and incubated on ice for 3 min. After washing with ATAC-RSB buffer (without NP40), nuclei were pelleted by centrifugation and incubated in 50 ml transposition mixture (25 ml 2 x TD buffer (Illumina, Ref 15027866), 2.5 ml transposase (Illumina, Ref 15027865), 16.5 ml PBS, 0.5 ml 1% digitonin, 0.5 ml 10% Tween-20, 5 ml H2O) at 37 °C for 30 min. Transposed DNA was purified using a Zymo DNA clean and concentrator-5 kit (Zymo research, Cat. No. D4014). Purified DNA was subjected to PCR amplification using the NEBNext® High-Fidelity 2 x PCR Master Mix (NEB, Cat. No. M0541s) system for first 5, and subsequent 3 cycles. Primer adapters were synthesized by IDT (Integrated DNA technology). Libraries were purified using a Zymo DNA clean and concentrator-5 kit, size-selected using AMPure XP beads, quantified using an Agilent 2100 Bioanalyzer Samples were pooled/sequenced using a NextSeq 500/550 High Output v2 kit using paired-end reads on a NextSeq 500 platform (Illumina).
GEO Accession
GSM4648814