title: GSM8261872: Pooled V(D)J Library; Mus musculus; OTHER
library_construction_protocol: Spleens were harvested at D+6 post-transplant (three mice pooled for group 1, four for group 2, and three for group 3). T cells from each mouse were incubated with APC-labeled CBir1 tetramer (NIH, above), washed, and labeled with TotalSeq C hashtag antibodies (C0301-C0310) along with TotalSeq C0987 (anti-AP). Following staining, T cells were sort-purified (Marilyn: CD45.1neg/CD45.2+/CD90.1+/CD90.2neg/CD4+/CD8neg; CBir1: CD45.1neg /CD45.2+/CD90.1neg/CD90.2+/CD4+/CD8neg) at a 4:1 ratio for cell capture. Library construction according to the 10x Genomics Chromium User Guide CG000424 (CG000424_ChromiumNextGEMSingleCell5_HTv2_CellSurfaceProtein_UserGuide_RevD.pdf), using Chromium Next GEM Single Cell 5'Kit (v2), Chromium Single Cell Mouse TCR Amplification Kit (v2), and Chromium Next GEM Chip K Single Cell Kit.