RNA-seq of Cuiguan peel at 0h control, biological replicate 1
Eukaryotic mRNA with polyA tails was enriched using magnetic beads coated with Oligo(dT), and then the mRNA was fragmented using fragmentation buffer. Using the fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA was synthesized in the M-MuLV reverse transcriptase system. Subsequently, the RNA strand was degraded by RNaseH, and the second strand of cDNA was synthesized using DNA polymerase I and dNTPs. The purified double-stranded cDNA was then subjected to end repair, A-tail addition, and ligation of sequencing adapters. The cDNA was then screened for fragments around 200bp using AMPure XP beads, and the selected cDNA was PCR amplified and further purified using AMPure XP beads to obtain the final library. The library construction principle is shown below.