GSM7678315: CD4 CD33 CAR T cells from AML patient Donor 11b, Day 14; 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. 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 NextSeq500 at a sequence depth of 10 million reads per sample.