Illumina HiSeq 4000 paired end sequencing; larval specimens of Xenocatantops brachycerus
Larval specimens, adult female specimens and adult male specimens (each specimens have 5 replicates) of X. brachycerus were collected in Xi'an, China, on September 12, 2015. The guts of the specimens were removed, and the remaining tissue was immediately frozen in liquid nitrogen. Total RNA from the three specimens of X. brachycerus were extracted individully using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. The purity, concentration, and integrity of the extracted RNA samples were then checked via gel electrophoresis using a NanoDrop 2000 (Thermo Scientific, Wilmington, DE, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, Mississauga, ON, Canada), no sign of degradation was found. The RNA samples were sent to BGI (Shenzhen, China) for RNA sequencing. Briefly, first-strand cDNA was synthesized from mRNA fragments using random hexamer primers and reverse transcriptase (Invitrogen, Carlsbad, CA, USA), and second-strand cDNA was synthesized using RNase H (Invitrogen, Carlsbad, CA, USA) and DNA polymerase I (New England BioLabs). The cDNA fragments were then ligated to adapter-containing primer sequences, and the products were amplified by PCR to generate the final cDNA library.
CNGB
CNX0000659