GSM4998319: Regressing3-TIL-VDJ; Mus musculus; RNA-Seq
(1) A223 tumors were removed from mice, digested using liberase into a single-cell suspension, and flow-sorted for CD8 T cells. (2) RNA and DNA were isolated from A223 cells grown in vitro (1) Single-cell gene expression libraries were prepared using Chromium Single Cell 5' Library & Gel Bead Kit (#1000014) or Chromium Single Cell 3' Library & Gel Bead Kit (#1000092) according to the manufacturer's instructions. 5' libraries were split in half, one for sequencing and another half was enriched for TCR sequencing using the Chromium Single Cell V(D)J Enrichment Kit (Mouse T Cell). (2) RNA: Libraries were prepared for sequencing using standard Illumina protocols. (2) DNA: Qualified genomic DNA samples were randomly sheared by the ultrasonic highperformance sample processing system (Covaris) into fragments with a main peak of about 200bp-300bp. Subsequently, repaired the end of DNA fragment, added nucleotide A at 3' end, and ligated adapters at both ends. The library ligated with adapters were linearly amplified (LM-PCR) to produce a hybrid library. Took an appropriate amount of hybrid library for capture and enrichment with exon chip, and eluted the unenriched fragments followed by amplification. The amplified products were subjected to QPCR quality control, qualified products were for sequencing. (1) Single-cell RNA sequencing (2) RNA: bulk RNA sequencing (2) DNA: Whole-exome sequencing