cDNA Cloning from Purified dsRNA

Purified dsRNA was used as a template for cDNA cloning. The cDNA library was constructed using the methods described by Zhong (Zhong et al., 2016 (link)). Random primers (5’-GCCGGAGCTCTGCAGAATTCNNNNNN-3’) and specific primers (5’-GCCGGAGCTCTGCAGAATTC-3’) were used for reverse transcription and PCR amplification. The intermediate sequences were amplified with specific primers designed according to the obtained sequences, and the 5’ and 3’ terminal sequences were obtained using adaptor-ligated methods as previously reported (Xie et al., 2011 (link); Zhong et al., 2016 (link)). All amplicons were cloned into the pMD18-T vector (Takara, Dalian, China) and transformed into Escherichia coli DH5α (TransGen Biotech, Beijing, China). Positive clones were selected for Sanger sequencing, with each base pair sequenced at least three times.