GSM8582023: D68-30-RNAseq; Homo sapiens; RNA-Seq
Total RNA from T cell clones was isolated from sorted T cells using the RNeasy Mini Kit (Qiagen). Cells in RLT buffer were vortexed for 1 minute to homogenize the lysate prior to adding 100uL of 80% ethanol, mRNA was then isolated following the manufacturer's instructions, and mRNA concentration was quantified using a NanoDrop™ 2000/2000c Spectrophotometer (ThermoFisher). Bulk RNA-seq libraries were prepared using NEBNext Ultra II RNA Library Prep (Illumina). Libraries (i.e., clones) were sequenced to an average depth of 30,000,000 reads using an Illumina HiSeq 4000 instrument or equivalent, with > 90% of bases being >Q30.