RNA-Seq of Baphicacanthus cusia (Nees) Bremek£ºLeaf
After total RNA was extracted, eukaryotic mRNA was enriched by Oligo(dT) beads, while prokaryoticmRNA was enriched by removing rRNA by Ribo-ZeroTM Magnetic Kit (Epicentre). Then the enrichedmRNA was fragmented into short fragments using fragmentation buffer and reverse transcripted intocDNA with random primers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTPand buffer. Then the cDNA fragments were purified with QiaQuick PCR extraction kit, end repaired,poly(A) added, and ligated to Illumina sequencing adapters. The ligation products were size selected byagarose gel electrophoresis, PCR amplified, and sequenced using Illumina HiSeqTM 4000 by Gene DenovoBiotechnology Co. (Guangzhou, China)