title: GSM5652385: CD4+ polyclonal CD1a-lysylPG-reactive T cell lines-donor 921a-unstimulated 3-921a-AdJ-unstim; Homo sapiens; RNA-Seq
library_construction_protocol: Reverse transcription was performed by adding 7.5 μl of RT mix to each well [containing: 40U Maxima H- (Thermo Fisher Scientific), 2X Maxima H- Buffer (Thermo Fisher Scientific), 4U SUPERaseIN (Thermo Fisher Scientific), 15% polyethylene glycol, 2µM Template Switching Oligo (Integrated DNA Technologies)]. Further sample processing and library preparation was performed by the Single Cell Analysis Core of Columbia University according to published protocol. For bulk RNAseq, RNA sequencing library preparation used the NEBNext Ultra RNA Library Prep Kit for Illumina by following manufacturer's recommendations (NEB, Ipswich, MA, USA). The samples were sequenced using a Illumina HiSeq 2x150 Paired End (PE) configuration. The single-cell RNA libraries were prepared for sequencing using Illumina NextSeq 500 using the 75-cycle High Outpit Kit [read lengths 26(R1) × 8(i) × 58(R2)].