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A method for species identification of venomous snakes has been developed on the basis of amplification
of a specific part of the mitochondrial cytochrome b gene using PCR. To distinguish among venomous
species, the obtained 1,144 bp PCR products were cut with different restriction endonuclease, Alu I and
Hinf I, resulting in species-specific restriction fragment length polymorphism (RFLP). Each enzyme
generated different-sized fragments which specific to the six neurotoxic and six hematotoxic snake species
in Thailand. However, identical patterns were found between Daboia siamensis and Trimeresurus
popeorum among hematotoxic snakes when Alu I has been used. These results could be resolved by using
additional enzyme such as Hinf I due to no cross reaction between the species was detected in their
restriction patterns. The PCR-RFLP developed here is simple, rapid, reliable and reproducible; hence it
can be routinely applied for snake species identification, essential for conservation and management of
endangered snake species in Thailand.
Keywords: PCR-RFLP, cytochrome b, neurotoxic, and hematotoxic snakes.