GSM9764234: A100_3; Homo sapiens; RNA-Seq
Total RNA was extracted using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. RNA quality and integrity were assessed by agarose gel electrophoresis and an Agilent 2100 Bioanalyzer system. RNA sequencing libraries were constructed using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina following the manufacturer's protocol. Briefly, mRNA was enriched using poly-T oligo-attached magnetic beads, fragmented, and reverse-transcribed to synthesize first- and second-strand cDNA. After end repair, adaptor ligation, and PCR amplification, libraries with insert sizes of approximately 250–300 bp were generated and sequenced on an Illumina NovaSeq platform to obtain paired-end 150-bp reads.