RNA sequencing of Oryza sativa  (japonica) roots
After enriching eukaryotic mRNA with polyA tails using magnetic beads with Oligo(dT), the mRNA was fragmented with buffer. Using the fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA was synthesized in an M-MuLV reverse transcriptase system. Subsequently, the RNA strand was degraded with RNaseH, and the second strand of cDNA was synthesized in a DNA polymerase I system using dNTPs as raw materials. The purified double-stranded cDNA was subjected to end repair, addition of an A tail, and ligation of sequencing adapters. AMPure XP beads were used to select cDNA of approximately 200 bp, followed by PCR amplification and purification of the PCR products with AMPure XP beads. The final library was obtained.