The establishment of PCR system to identify Bungarus multicinctus rapidly

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus.  Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus.  On the basis of the sequence data, a pair of highly specialized primers was designed.  The templates were extracted by the DNA purification system.  Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized.  The modified PCR program consisted of an initial denaturation step at 95 ℃ for 5 min, followed by 30 cycles of 95 ℃ for 30 s and 55 ℃ for 45 s and a final extension at 72 ℃ for 5 min.  Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours.  The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.