PCR-RFLP analysis of the internal transcribed spacer (ITS) region for identification of 3 clam species

Restriction site analysis of the internal transcribed spacer (ITS) region a mplified by the polymerase chain reaction (PCR) was used for the specific i dentification of 3 clam species: Ruditapes decussatus (grooved carpet shell ), Venerupis pullastra (pullet carpet shell), and Ruditapes philippinarum ( Japanese carpet shell). PCR amplification using pruners based on nucleotide sequences of Mytilus ITS regions produced a fragment of 1195 bp in R. decu ssatus, 1074 bp in V pullastra, and 1188 bp in R. philippinarum. Digestion of the PCR products with endonucleases Hinf I and Rsa I, followed by agaros e gel electrophoresis of the digested products, yielded specific restrictio n profiles that enabled direct visual identification of the species analyze d.