GSM5513773: Cd45+ Day_3_T; Mus musculus; RNA-Seq
After washing and resuspending in an appropriate volume of cold PBS, purified Cd45+ cells were processed in the Chromium single cell gene expression platform (10x Genomics). In brief, Single Cell 3′ Reagent Kit v2 (10X Genomics) was used to generate droplets encapsulating single cell and barcoded bead. Following lysis, reverse transcription was performed in a thermal cycler (ProFlexTM PCR). cDNA was purified with Dynabeads (provided by 10x) and amplified for 10 or 14 cycles (ProFlexTM PCR). The amplified-cDNA was fragmented, ligated with adapter and sample index, and selected with SPRI beads (Beckman). The constructed library was sequenced on the NovaSeq 6000 Illumina sequencing platform.