title: GSM2356790: invitro NFAT CARIT rep2, ATAC-seq; Mus musculus; OTHER
library_construction_protocol: ATAC-seq library preparations were performed essentially as described (Buenrostro et al., 2013), with minor modifications. Briefly, 50,000 cells were washed once in PBS before incubation for 30 minutes in lysis buffer (10 mM Tris pH 7.5, 10 mM NaCl, 3mM MgCl2, 0.1% NP-40). For one replicate of memory cells and d35 KLRG1+ cells, 10,000 cells were used. After lysis, cells were resuspended in 50 ul 1x TD Buffer containing 2.5 ul Nextera enzyme (Illumina, San Diego, CA).  Transposition reactions were incubated for 30 min at 37°C before purification with QiaQuick MinElute columns (Qiagen, Valencia, CA). Purified DNA was amplified by PCR using Kapa Hi-Fi real-time library amplification kit (Kapa, Wilmington, Massachusetts) with 10-12 cycles, according to the manufacturer's instructions with barcoded primers, as described previously (Buenrostro et al., 2013).  Amplified libraries were purified with QiaQuick MinElute columns (Qiagen, Valencia, CA) and quantitated with Kapa real-time library quantification kit (Kapa, Wilmington, Massachusetts). Paired-end sequencing was performed with the rapid run protocol with an Illumina HiSeq 2500 (Illumina, San Diego, CA) with 50 cycles in each direction.