GSM7508112: D-Red+; Mus musculus; RNA-Seq
Single cell suspensions were generated in 1 % v/v FCS in PBS by meshing fresly isolated lymph nodes, incubated with fluorochrome labelled antibodies for 20 minutes on ice and sorted with a BD FACSAria Fusion sorter. Library construction was performed according to manufacturer's instructions (Single Cell 5' R2-only protocol, 10x Genomics). Briefly, single cells were encapsulated into droplets with the ChromiumTM Controller (10x Genomics) and processed following manufacturer's specifications. Transcripts captured in all the cells encapsulated with a bead were uniquely barcoded using a combination of a 16 bp 10x Barcode and a 10 bp unique molecular identifier (UMI). cDNA libraries were generated using the Chromium™ Next GEM Single Cell 5' Reagent Kit v2 (10x Genomics) following the detailed protocol provided by the manufacturer.