title: GSM6943499: Ex-Vivo human T-cells, 80BB scRNAseq; Homo sapiens; RNA-Seq
library_name: GSM6943499
library_construction_protocol: For single-cell RNA sequencing analysis, 5 Mice per group were injected with NALM6 tumour and treated with 1?×?1e5 or 5?×?1e4 19-HIT+ T cells per mouse on day 4 post tumour injection. On day 9 post T cell injection, bone marrow cells were isolated.Cells from mice within one condition were pooled and labelled with PE mouse anti-human CD45 (2D1, Biolegend) and TotalSeq™-B0251 anti-human Hashtag 1 Antibody (Biolegend) in 19-HIT treated mice or TotalSeq™-B0254 anti-human Hashtag 4 Antibody (Biolegend) in 19-HIT80BB treated mice.  Cells were FACS sorted for CD45+ GFP- cells (human T cells, excluding NALM6).  Cells from different conditions were pooled together at a balanced ratio and submitted for 10X 3' RNA sequencing. Cells were stained with Trypan blue and Countess II Automated Cell Counter (ThermoFisher) was used to assess both cell number and viability. Following QC, the single cell suspension was loaded onto Chromium Next GEM Chip G (10X Genomics PN 1000120) and GEM generation, cDNA synthesis, cDNA amplification, and library preparation of 15-21,000 cells proceeded using the Next GEM Single Cell 3' Kit v3.1 (10X Genomics PN 1000268) according to the manufacturer's protocol. cDNA amplification included 11 cycles and 10 µL of the material was used to prepare sequencing libraries with 8-10 cycles of PCR.