GSM5904519: Day 7 Treg; Homo sapiens; ATAC-seq
ATAC-seq was performed from 50,000 Treg cells per a previously published protocol (Buenrostro, Nature Methods, 2013). Briefly, nuclei were prepared by washing cells with PBS, followed by lysis using cold lysis buffer. Nuclei were spun and the pellet was then resuspended in the transposase reaction mix for 30 min at 37 °C. The samples were then purified using a Qiagen Minelute kit. Amplified libraries were purified, and the sizes of library DNA were determined by the Fragment Analyzer (Advanced Analytical Technologies, IA) using the High Sensitivity NGS Fragment Analysis Kit (Cat # DNF-486). The enrichment of accessible region was determined by the fold difference between positive and negative genomic loci using real-time PCR. The libraries were sequenced to 51 base pairs from both ends on an Illumina HiSeq instrument in the Mayo Clinic Center for Individualized Medicine Medical Genomics Facility.