Kaloula pulchra Transcriptome or Gene expression. Kaloula pulchra

Total RNA was extracted using TRIZOL reagent (Takara Bio Inc., Otsu, Japan). The degradation and purity of total RNA were examined through 1% agarose electrophoresis analysis and the NanoPhotometer spectrophotometer (IMPLEN, CA, USA), respectively. The integrity was also further demonstrated with the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). The cDNA library was constructed and sequenced by Novogene (Beijing, China). In brief, poly-Toligo-attached magnetic bead was used to purify the mRNA from total RNA which was subsequently fragmented with divalent cations under enhanced temperature. Then, the fragmented mRNA was used as the template for the synthesis of the first strand of cDNA with M-MuLV Reverse Transcriptase and a random hexamer primer.