Transcriptomic analysis of Stevia rebaudiana MS007
After the QC, mRNA from Stevia rebaudiana was enriched using oigo(dT) beads. The mRNA from  Stevia rebaudiana  was fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamers and reverse transcriptase. After first-strand synthesis, a custom second-strand synthesis buffer (Illumina) is added, with dNTPs(the dTTP is replaced by dUTP), RNase Hand Escherichia coli polymerase I to generate the second strand by nick-translation and AMPure XP beads is used to purify the cDNA, then the second strand is degraded by USER Enzyme. The final cDNA library is ready after a round of purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR enrichment. Library concentration was first quantified using a Qubit 2.0 fluorometer (Life Technologies), and then diluted to 1 ng/l before checking insert size on an Agilent 2100 and quantifying togreater accuracy by quantitative PCR (Q-PCR) (library activity >2 nM).