GSM9153277: 2269_V1+1 (tempus); Homo sapiens; RNA-Seq
Whole blood RNA was extracted from Tempus tubes in batches of 12-24 samples, with a mixture of vaccination groups and timepoints, using the Tempus Spin RNA Isolation Kit (Thermo Fisher Scientific). Samples were thawed, diluted in a 50 ml conical with 1× PBS, vortexed for 30 s, then centrifuged (3000 × g, 30 min, 4°C) to pellet RNA. Supernatants were removed and RNA pellets resuspended in RNA Purification Resuspension Solution, then transferred to purification filters for microcentrifuge RNA elution. RNA quantity and integrity was assessed using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific) and Agilent TapeStation and submitted to the Oxford Genomics Centre (Centre for Human Genetics, University of Oxford) for library generation and sequencing (Illumina NovaSeq 6000, 150 base pair paired-end reads). NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, #E7760) with Globin & rRNA Depletion (New England Biolabs, #E7750).