GSM8775010: scRNA-seq, Adult CD8+ Recent Thymic Emmigrant T cells, Rep1; Mus musculus; RNA-Seq
Murine cells were isolated from spleen two weeks post timestamp. Single cell suspensions were obtained by manual dissociation and filtration through a 40uM strainer. CD8+ T cells were isolated by positive magnetic separation using anti-CD8a microbeads. Live Va2+CD8+ cells were sorted into 0.04% BSA in PBS. Cells were loaded in the Chromium instrument (10X Genomics) for the formation of gel bead-in-emulsions (GEMs) targeting 5-10k cells per sample. Single-cell RNA-seq and TCR libraries were prepared using the Chromium Single Cell V(D)J Reagent Kit (v1.1 Chemistry) (10X Genomics) by the BRC Genomics Facility following the manufacturer's protocol. Libraries  were sequenced on a NovaSeq 6000  (Illumina) to an average depth of >150M reads (scRNA-seq) and >15M reads (scTCR).