GSM9821727: Whole blood, crystalloid, RIE-14, T2; Homo sapiens; RNA-Seq
Whole blood collected into Tempus Blood RNA Tubes (Thermo Fisher #4342792). Total RNA extracted with the MagMAX for Stabilised Blood Tubes RNA Isolation Kit (Thermo Fisher #4451894). RNA integrity assessed on an Agilent Fragment Analyser (#5300, Standard Sensitivity RNA Analysis Kit DNF-471-0500; mean RQN 3.9); RNA and DNA quantified by Qubit (RNA broad-range #Q10210; dsDNA HS #Q32854). Samples with >20% DNA contamination were excluded; 69 of 76 samples proceeded to library preparation. Libraries prepared from 250 ng total RNA using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760) with the NEBNext Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7550X). Stranded (dUTP second-strand) protocol; 11 cycles of PCR; barcoded with NEBNext Multiplex Oligos for Illumina Unique Dual Index Primer Pairs (NEB #E6442L).