RNA-Seq of tea leaves in 3h under high temperature stress
The integrity of extracted RNA was detected by Bioanalyzer 2200 (Agilent) and RNA integrity number larger than 8 is acceptable for complementary DNA (cDNA) library construction. The cDNA libraries were constructed using the TruSeq Stranded mRNA Library Prep Kit (Illumina, Inc.) . mRNA was purified by oligo (dT) magnetic beads, and fragmented into 200-500 bp length using divalent cations at 94 for 5 min. The cleaved RNA fragments were used for First-strand cDNA synthesis through reverse-transcriptase and random primers. Second-strand cDNA was synthesized by cDNA and dNTP mix. The products were purified and enriched by PCR to obtain final cDNA libraries and sequenced by HiseqX11 platform