GSM8490311: V_A; Mus musculus; RNA-Seq
Subcutaneous tumor tissues were dissociated at 37 ℃ with a shaking speed of 50 r.p.m for about 40 min.We repeatedly collected the dissociated cells at interval of 20 min to increase cell yield and viability. Cell suspensions were filtered using a 40 μm nylon cell strainer and red blood cells were removed by 1× Red Blood Cell Lysis buffer (Thermo Fisher Scientific, cat. no. 00-4333-57) . Dissociated cells were washed with 1× DPBS containing 2% FBS. Cells were stained with AO/PI, and viability was assessed using a Countstar Fluorescence Cell Analyzer (Aber Instruments) Sequencing libraries were prepared using randomly interrupted whole-transcriptome amplification products to enrich the 5' end of the transcripts linked with the cell barcode and UMI. Single-cell RNA libraries were prepared using the Chromium Single Cell 5'v2 Reagent (10x Genomics), and TCR/BCR transcripts were enriched with the Chromium Single Cell V(D)J Reagent kits (10x Genomics). Each sequencing library was generated with a unique sample index.