Libraries for sequencing were prepared using the Illumina TruSeq Stranded mRNA Library Prep kit by loading 50 ng of total RNA into the initial reaction; fragmentation and PCR steps were undertaken as per the manufacturer’s instructions.

Final library concentrations were determined using Qubit 2.0 fluorometer and pooled to a normalized input library.

Pools were sequenced using the Illumina NovaSeq 6000 Sequencing system to generate 150 bp paired-end reads with an average read depth of ~137 million paired-end reads per sample.