GSM9384234: 502_TNY9R23; Homo sapiens; RNA-Seq
Whole blood was collected in Tempus Blood RNA Tubes (Thermo Fisher Scientific), RNA was extracted using MagMax for Stabilized Blood Tubes RNA Isolation Kit (Thermo Fisher Scientific), followed by globin reduction using GlobinClear Human (Thermo Fisher Scientific). To generate sequencing libraries, total RNA (0.5 ng) was added to reaction buffer from the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Takara), and reverse transcription was performed followed by PCR amplification to generate full-length amplified cDNA. Sequencing libraries were constructed using the NexteraXT DNA sample preparation kit (Illumina) to generate Illumina-compatible barcoded libraries. Libraries were pooled and quantified using a Qubit Fluorometer (Life Technologies). Dual-index, single-read sequencing of pooled libraries was carried out on a HiSeq 2500 sequencer (Illumina) with 58-base reads, using HiSeq v4 Cluster and SBS kits (Illumina) with a target depth of 5 million reads per sample.