GSM5931291: D18_Low_CD8_d0_S26_R1_001 [RNA-seq]; Homo sapiens; RNA-Seq
RNA-seq was performed following MARS-seq protocol adapted for bulk RNA-seq with minor modifications. Briefly, 7000 to 10000 cells were sorted in 100µl of Lysis/Binding Buffer (Ambion), vortexed and stored at -80ºC until further processing. Poly-A RNA was reverse-transcribed using poly-dT oligos carrying a 7 nt-index. Pooled samples were subjected to linear amplification by IVT. Resulting aRNA was fragmented and dephosphorylated. Ligation of partial Illumina adaptor sequences was followed by a second reverse-transcription reaction. Full Illumina adaptor sequences were added during final library amplification. RNA-seq libraries quantification was done with Qubit 3.0 Fluorometer (Life Technologies) and size profiles were examined using Agilent's 4200 TapeStation System. Libraries were sequenced in an Illumina NextSeq 500 at a sequence depth of 10 million reads per sample. MARS-seq (Illumina)