GSM8524622: CD3 T cells, in vitro cell culture Day2, 4pm replicate3; Homo sapiens; RNA-Seq
Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA,USA) according to the manufacturer's protocol.After total RNA was extracted, eukaryotic mRNA was enriched with oligo (dT) beads. The enriched mRNA was subsequently fragmented into short fragments via fragmentation buffer and reverse transcribed into cDNA via the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #7530, New England Biolabs, Ipswich, MA, USA). The purified double-stranded cDNA fragments were end repaired, poly(A) was added, and the fragments were ligated to Illumina sequencing adapters. The ligation reaction mixture was purified with AMPure XP beads (1.0X), and polymerase chain reaction (PCR) amplification was performed.