GSM6161300: Lymph_node_endogenous_RNA; Mus musculus; RNA-Seq
Sorted T cells were stained with Trypan blue and the Countess II Automated Cell Counter (ThermoFisher) was used to assess both cell number and viability. Following QC, the single cell suspension was loaded onto Chromium Next GEM Chip K (10X Genomics PN 1000286) and GEM generation, cDNA synthesis, abd cDNA amplification. Library preparation of 8,100 cells proceeded using the Chromium Next GEM Single Cell 5' Kit v2 (10X Genomics PN 1000263) according to the manufacturer's protocol. cDNA amplification included 13 cycles and 32ng of the material was used to prepare a sequencing library with 14 cycles of PCR. The indexed library was pooled equimolar and sequenced on a NovaSeq 6000 in a PE28/91 run using the NovaSeq 6000 SP Reagent Kit (100 cycles) (Illumina). 228 million paired reads were generated for the sample.