RNA-Seq of Spodoptera litura: midgut of 5th instar larvea feeding on Brassica juncea for 6h
Firstly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads.Then mRNA was break into short pieces with fragmentation buffer. First strand cDNA was synthesized using random hexamer primer. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. The overhangs of DNA fragments were converted into blunt ends. After adenylation of 3ĄŻ ends of DNA fragments, sequence adaptor was added. 150~200 bp length  cDNA fragments were purified with AMPure XP beads. Then PCR was performed and products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system.