GSM6644201: whole blood, subject a4a38a22, sample time point T4, library prep Plate_17; Homo sapiens; RNA-Seq
Frozen blood samples were thawed and total RNA was extracted from the samples using a modification of the MagMax protocol for Stabilized Blood Tubes RNA Isolation Kit (Thermo Fisher, #4451893). Samples that yielded sufficient RNA (>50 ng) were barcoded and prepared for pooled whole transcriptome sequencing using the TruSeq Stranded Total RNA Library Prep Gold (Illumina, #20020599), which is designed to remove ribosomal, globin, and mitochondrial RNA. Libraries were amplified with 15 cycles of PCR, pooled, and sequenced on a NovaSeq 6000 (Illumina) using Sprime flow cells with 100 base pair paired-end reads, targeting a mean of 50 million read pairs per sample. For a minority of samples in which the first extraction failed (N=24), RNA was re-extracted from the supernatant saved from the first centrifugation pellet. The extraction protocol was repeated starting with the second wash step after re-pelleting the RNA.