RNA-Seq of rice infected with Aphelenchoides besseyi
Eukaryotic mRNA was enriched by ployA tail binding of A-T complementary pairing with ployA tail of mRNA using magnetic beads with Oligo (dT). Then fragmentation buffer was added to break the mRNA into short fragments. A single strand of cDNA was synthesized with random hexamers using the template of the mRNA. Then the buffer, dNTPs and DNA polymerase I were added to synthesize the double-stranded cDNA. Then the double-stranded cDNA was purified by AMPure XP beads. The purified double-stranded cDNA was repaired at the end, A-tail was added and sequencing junction was connected, then the fragment size was selected by AMPure XP beads, and the final cDNA library was enriched by PCR