Authentication of nine snapper species by single-strand conformation polymorphism (SSCP) and forensically informative nucleotide sequencing (FINS) methods

PCR-SSCP and PCR-FINS methods were developed to authenticate nine species of snapper. A highly variable mitochondrial D-loop region was chosen as a target region and three new primers viz., one forward (Fish-DL-F) and two reverse primers (Fish-DL-R and DLFR) were designed. PCR-SSCP was first targeted to amplify 515 bp D-loop fragment using Fish-DL-F/Fish-DL-R primers, but it could not differentiate all the species. Another reverse primer (DLFR) could amplify a 360 bp region, which had differentiated all the species. A FINS method was also developed for snapper authentication using the same D-loop region of 515 bp. The developed FINS was validated with 20 market snapper samples that showed 100% specificity. This study helped to generate nucleotide sequences of D-loop region for 6 species of snapper for the first time for deposition in GenBank. Hence, to authenticate the snapper species, the developed PCR-SSCP and FINS were more specific and reliable.