Participant 8, skin sample taken 6 hours after surgical incision
Poly-A selected stranded mRNA libraries were constructed from 1 µg total using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep for Illumina with Sample Purification Beads (NEB #E7765, New England Biolabs, Ipswich, MA). Amplification was performed using 8 cycles to minimize the risk of over-amplification. Unique dual-indexed barcode adapters were applied to each library. Libraries were pooled in an equimolar ratio for sequencing. The pooled libraries were sequenced on an S4 flow cell on a NovaSeq 6000 (Illumina, San Diego, CA) using v1.5 chemistry to achieve a minimum of 60 million 150 base read pairs. The data was processed using RTA version 3.4.4.