title: GSM6599283: P14, IL2Rb ortho; Mus musculus; RNA-Seq
library_construction_protocol: Sorted samples were topped with cold PBS 0.04% BSA, centrifuge for 5' 350g at 4oC, washed two times in cold PBS and resuspended in 50-100ul of cold PBS. Samples were counted, down-sampled and equivalent number of cells (6300) between samples were loaded into the Chip (Chromium Next GEM Chip G) of a Chromium Next GEM Single Cell 3' Reagent Kit v3.1 (Dual Index, 10x Genomics) and run onto a Chromium Controller Samples were then processed according to manufacturer's protocol. cDNA libraries were prepared using the Dual Index TT Set A (10x Genomics) and the number of indexing PCR cycles was adjusted to the cDNA input ,Libraries were quantified by qPCR using a KAPA Library Quant Kit (KAPA Biosystems). Normalized libraries were pooled (2.5nM), loaded onto a NovaSeq 6000 SP Reagent Kit (100 cycles, Illumina) for a final concentration of 450pM and paired-end sequencing was performed on a NovaSeq 6000 (Illumina).