Sample information curated by ChIP-Atlas

Antigen

Antigen Class
ATAC-Seq
Antigen
ATAC-Seq

Cell type

Cell type Class
Blood
Cell type
CD8+ T cells
NA
NA

Attributes by original data submitter

Sample

source_name
tumor-infiltrating lymphocytes
strain
C57BL/6J CD45.1 OT-I
cell type
CD8+ T cells
treatment
NA

Sequenced DNA Library

library_strategy
ATAC-seq
library_source
GENOMIC
library_selection
other
library_construction_protocol
Cell suspensions were prepared from tumors after enzyme digestion. For sorting of adoptively transferred OT-I and P14 cells, CD8+ T cells were positively selected from tumors using the Dynabeads FlowComp Mouse CD8 selection kit (Thermo Fisher). To ensure adequate cell yield, samples from multiple mice belonging to the same group were pooled prior to enrichment. Enriched CD8+ T cells were then stained with anti-CD8a (53-6.7), anti-CD45.1 (A20), anti-Thy1.1 (OX-7), anti-PD-1 (29F.1A12), anti-Tim-3 (RMT3-23) and Fixable Viability Dye eFluor780. Tumor infiltrating lymphocytes were isolated as live CD8+CD45.1+ (OT-I) or CD8+Thy1.1+ (P14) on BD FACSARIA III or FUSION sorters. ATAC-seq was performed according to Buenrostro et al. (Nature Methods 2013) with minor modifications: 30,000 to 50,000 sorted CD8+ T cells cells were lysed to extract nuclei. Nuclei were resuspended in 50 ul 1xTD Buffer containing 2.5 μl transposase (Nextera, Illumina). The transposase reaction was conducted for 30 minutes at 37°C, with mild shaking. Library amplification and barcoding were performed with the library amplification kit (KAPA Biosystems) using Index Primers, designed according to Buenrostro et al. (Nature Methods 2013) at a final concentration of 500 nM. PCR reaction was conducted for 10-11 cycles. Library purification was performed with the MinElute PCR Purification Kit (Qiagen) and library size distribution was assessed using the Bioanalyzer High Sensitivity DNA Kit (Agilent). ATAC-seq libraries were quantified prior to pooling and sequencing using the real-time library quantification kit (KAPA Biosystems).

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
36244531
Reads aligned (%)
97.1
Duplicates removed (%)
31.0
Number of peaks
33045 (qval < 1E-05)

mm9

Number of total reads
36244531
Reads aligned (%)
96.9
Duplicates removed (%)
31.1
Number of peaks
33046 (qval < 1E-05)

Base call quality data from DBCLS SRA