Cloning of Full-Length cDNA of Teleost Corticotropin-Releasing Hormone Precursor by Improved Inverse PCR

Abstract

We designed a new inverse PCR protocol combined with switching mechanism at 5′ end of RNA transcript (SMART) technology, and applied it to the cloning of teleost corticotropin-releasing hormone precursor cDNA. Due to the advantages of both techniques, this method can efficiently amplify the complete 5′- and 3′-ends of cDNA in a single reaction, and might prove to be an alternative to the conventional rapid amplification of cDNA ends (RACE) approaches.