RNA-Seq of Liriomyza sativae: adult
A total of 10 individuals, which were raised in laboratory for ten generations, were pooled and used for RNA extraction based on TRIzol reagent (Invitrogen, USA). Using high-quality extracted RNA, mRNA was enriched using magnetic beads with Oligo (dT) (Illumina, Inc). Subsequently, fragmentation buffer was used to break mRNA into short fragments. Using mRNA as a template, single-strand cDNA was synthesized using random hexamers as primers. Double-stranded cDNAs was then synthesized by adding a mixture of buffer, dNTPs, DNA polymerase I and RNase H, and purified using AMPure XP beads to isolate end-repaired and a-tailed products. PCR amplification and repurification using AMPure XP were performed. Finally, library quality was assessed using Agilent Bioanalyzer 2100 system. After the library was qualified, Illumina HiSeq sequencing was performed to generate paired-end reads.