GSM5851362: CD4SP&CD25pos&FOXP3lo&Treg&CD73pos&CD4CD8dp [TCR]; Mus musculus; RNA-Seq
Mechanically dissociated thymus depleted of CD8 via negative magnetic enrichment (double postive thymocytes were captured from non-enriched whole thymocytes) was sorted via flow cytometry of a BD FACSAria II to isolate each input cell population. Cells were sorted into 1xPBS + 50% FBS prior to hashtag staining then resuspension in 1xPBS + 5% FBS @ 3000 cells/uL for single cell library preparation. The 10x Genomics Chromium Next GEM Single Cell 5' Kit (v2) was used to generate three libraries: (1) mRNA transcript expression (RNA), (2) mouse-specific hastag oligos (HTO) and (3) sequencing of single T-cell receptor sequence (TCR) sequence (specifically enriched for TCR alpha sequence).