title: RNA-Seq of Crocus sativus
design: There are two main ways to obtain mRNA: one is to utilize the structural feature that most of the eukaryotic mRNAs have polyA tails, and enrich the mRNAs with polyA tails by Oligo(dT) magnetic beads; the other is to remove the ribosomal RNA from the total RNA, so as to get the mRNAs. subsequently, add fragmentation buffer to interrupt the RNAs into The first strand cDNA was synthesized by using short fragment RNA as a template with six-base random primers (random hexamers), and then two-stranded cDNA was synthesized by adding buffer, dNTPs (dTTP, dATP, dGTP, and dCTP), and DNA polymerase I. Subsequently, double-stranded cDNA was purified by using DNA-purifying magnetic beads. The purified double-stranded cDNA was then subjected to end repair, A-tailed and ligated to sequencing junctions, followed by fragment size selection using DNA purification magnetic beads, and finally PCR enrichment to obtain the final cDNA library.