title: GSM4555171: Tumor specific T cell, WT3; Mus musculus; ATAC-seq
library_construction_protocol: A total of 1 × 10e5 B16-F10 cells were injected subcutaneously into 6 to ~10-week-old female C57BL/6J mice (six mice per group). On day 21, mice were euthanized and subcutaneous tumors were digested with 1 mg/ml collagenase D supplemented with 10 U/ml DNase I. OVA-specific CD8+ TILs were sorted using a BD FACS Aria cell sorter, and gene profiling, ATAC-seq library construction and high-throughput sequencing were performed on approximately 50,000 sorted cells. Libraries were prepared according to Vazyme's instructions accompanying the DNA Sample Kit (TD501). Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3' end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina Hiseq 4000 instrument following the manufacturer's protocols.
attributes.tag: GEO Accession
attributes.value: GSM4555171