title: GSM7865658: H2-O KO CD4 Tcell, Replicate 3; Mus musculus; RNA-Seq
library_construction_protocol: Total splenocytes were isolated from H2-O WT and H2-O KO spleens using manual tissue disruption and 70 um filtration. Red blook cells (RBCs) were lysed using ACK (4mL, 3mins on ice). Cells were washed 2 times with RPMI + 10% FBS to remove ACK buffer. Recovered cells were surface stained with anti-CD3, -CD4,-B220, and -Nk1.1 for 30mins at 4 degree. PI was added right before sorting for dead cell exclusion. Sorted cells were collected into HBSS + 10% FBS and submitted for scRNA-Seq analysis. The samples were prepared using the 10x Genomics Chromium Next GEM Single Cell 5' Library and Gel Bead Kit v1.1, Chromium Next GEM Chip and Dual Index Kit TN Set A. They were run on the Illumina NovaSeq6000 with a run configuration of 28bp x 10bp x 10bp x 91bp.