GSM5444429: CD8_neg_neg_NYBC230_run2_58; Homo sapiens; RNA-Seq
RNA was isolated from sorted cell populations using TRIzol (Invitrogen). After RiboGreen quantification and quality control by Agilent BioAnalyzer, 0.27-2.0ng total RNA with RNA integrity numbers ranging from 3.3 to 10 underwent amplification using the SMART-Seq v4 Ultra Low Input RNA Kit (Clonetech catalog # 63488), with 12 cycles of amplification. Subsequently, 5.7-7.0ng of amplified cDNA was used to prepare libraries with the KAPA Hyper Prep Kit (Kapa Biosystems KK8504) using 8 cycles of PCR. Samples were barcoded and run on a NovaSeq 6000 in a PE50 run, using the NovaSeq 6000 S1 or S2 Reagent Kit (100 Cycles) (Illumina).  An average of 71 million paired reads were generated per sample and the percent of mRNA bases per sample ranged from 29% to 65%.