RNA-Seq of cucumis sativus: temporal study
After the QC procedures, mRNA was enriched using oligo(dT) beads and library was constructed by Novogene. rRNA was removed using the Ribo-Zero kit that leaves the mRNA. First, the mRNA was fragmented randomly by adding fragmentation buffer, then the cDNA was synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I were added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library was completed through size selection and PCR enrichment.