RNA-seq of Kmeria septentrionalis: leaf
Total RNA was extracted from leaf samples using the mirVana miRNA Isolation Kit (Thermo Fisher Scientific) according to the manufacturer's instructions. RNA purity and concentration were assessed using a NanoDrop spectrophotometer (Thermo Scientific) and agarose gel electrophoresis. Only high-quality RNA samples with A260/A280 ratios between 1.8-2.0 were used for subsequent analysis.mRNA was isolated from 50 g of total RNA using oligo(dT) beads from the Stranded mRNA Library Prep Kit. First-strand and second-strand cDNA were then synthesized. Overhangs were adenylated, and universal Illumina PCR adapters were ligated to the cDNA fragments. The prepared cDNA libraries were indexed and pooled in equimolar ratios. The pool was sequenced on the BGI system with paired-end 150bp reads at ~3Gb per library.