RNA-Seq of Phytolacca americana leaves
(1) The mRNA was enriched with Oligo (dT) magnetic beads; (2) The mRNA was randomly broken into short fragments with fragmentation buffer; (3) The first cDNA strand was synthesized using random hexamers primer, and then the second cDNA strand was synthesized using DNA polymerase I, dNTPs and RNase H. The double-strand cDNA was purified with AMPure XP beads; (4) The purified double-strand cDNA was performed with end reparation, adding A tail and ligation to the sequencing adaptors, and then AMPure XP beads were used for fragment size selection; (5) The purified cDNA template was enriched with PCR amplification.