RNA-seq of Rana sylvatica: juvenile dorsal skin
Whole body was stored in RNAlater (Sigma, cat. no. R0901) at -20 _ in the field, then at -80 _ upon receipt at the University of Waterloo. Body was removed from RNAlater, individual tissues dissected, flash-frozen in liquid nitrogen, and stored at -80 _ until RNA isolation. Total RNA was isolated using TRI Reagent (Sigma, cat. no. T9424). This sample additionally underwent ethanol precipitation prior to sequencing to improve quality ratios. Enrichment of mRNA and library preparation was performed by The Centre for Applied Genomics (TCAG; The Hospital for Sick Children, Toronto, Ontario, Canada) using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB), followed by paired-end 150 base pair sequencing on one lane of a NovaSeq 6000 S4 flowcell.