GSM9210838: gamma-delta T cells from healthy donor 1, untreated; Homo sapiens; RNA-Seq
Total RNA was extracted from PBMC-derived gamma-delta T cells using TRIzol reagent (Thermo Fisher Scientific) according to the manufacturer's protocol. RNA integrity was assessed using an Agilent 2100 Bioanalyzer, and only samples with RIN ≥ 7 were used for library preparation. mRNA was enriched from total RNA using oligo(dT)-attached magnetic beads. The enriched poly(A) RNA was fragmented using fragmentation buffer, and first-strand cDNA was synthesized with random hexamer primers, followed by second-strand synthesis to obtain double-stranded cDNA. The double-stranded DNA was end-repaired and 5′-phosphorylated, with a 3′-A overhang added. A bubble-shaped adapter with a 3′-T overhang was ligated to the fragments, and ligation products were PCR-amplified with specific primers. The PCR products were denatured into single strands, and single-stranded circular DNA libraries were generated using a bridging primer. Final libraries were quality-checked and sequenced once they passed QC