title: GSM9209474: Tem _P14, tumor, Rep1; Mus musculus; ATAC-seq
library_construction_protocol: To obtain P14 CD8 T cells in tumor, tumors were mechanically minced and digested using with 1 mg/mL collagenase I (Sigma, c0130) and mashed through filters. Then, lymphocytes were enriched using Percoll (GE Healthcare, 17-0891-09) density gradient. Next, lymphocytes were stained with flow cytometry antibodies specicifc for CD8, Va2, CD45.1, CD45.2, P14 cells were FACS-sorted. ATAC-seq library was prepared using Hyperactive ATAC-seq Library Prep Kit for Illumina (Vazyme, TD711). Briefly, sorted cells were lysed on ice and fragment DNA in waterbath at 37℃ for 30 min. ATAC DNA Extract Beads were used for DNA extraction of fragmented products and extracted DNA fragments were amplified in a PCR instrument. Finally, ATAC DNA Clean Beads were used for two-step fragment size selection of amplified products and quality and quantity assessments are conducted using methods such as bioanalyzer or Qubit. Finally, the libraries were sequenced using paired-end sequencing, with 150 nucleotides on each end, conducted in a single lane of a NovaSeq 6000 for each sample.