GSM4010397: 6-4_CD8; Homo sapiens; RNA-Seq
Cells were collected 5 days after transduction and CD4+ and CD8+ cells were isolated via flow cytometry assisted cell sorting (FACS) on a BD FACS Aria III. RNA was isolated using the RNeasy Mini Kit (Qiagen). mRNA sequencing libraries were prepared using the NuGEN Universal Plus mRNA-seq + UDI kit (NuGEN 9144-96). Briefly, 1ug of Human Primary T cell total RNA (Agilent TapeSataion RIN values between 9.7 - 10) was enriched for mRNA by oligo-dT bead capture, fragmented, reverserse transcribed into cDNA. cDNA was end-repaired and ligated with "adapter-dimer free" Unique Dual Index (UDI) adapters, strand selected, PCR amplified with 12 cycles of amplification followed by KAPA Pure Bead (KAPA KK8002) clean-up and sizing resulting in a 290bp median sized library. 100 cycle Paired end sequencing data was generated with an Illumina NovaSeq 6000 Sequencer using NovaSeq 6000 S1 Reagent Kit - 200 cycle (Illumina 20012864).