Total RNA was isolated from 10 larvae of dsTcAMPKα or dsEGFP group on the sixth day after injection using TRIzol Reagent (Invitrogen, USA), and digested by RNase-free DNase I (Takara, Dalian, China) to remove genomic DNA contaminants. 1. Enrichment of eukaryotic mRNA by magnetic beads with Oligo (DT). 2. The mRNA was randomly interrupted in fragmentation buffer. 3. The first cDNA strand was synthesized by random hexamers using mRNA as template. The second cDNA strand was synthesized by adding buffer, dNTPs, RNase H and DNA polymerase I. Purification of cDNA by ampure XP beads. 4. The purified double stranded cDNA was subjected to terminal repair, A-tail was added and sequenced. The fragment size was selected by ampure XP beads. 5. Finally, the cDNA library was obtained by PCR enrichment.
GEO Accession
GSM4709235