title: [The establishment of PCR system to identify Bungarus multicinctus rapidly].
abstract: 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 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C 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.
authors.affiliation: College of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
authors.forename: Jing-xue
authors.initials: JX
authors.lastname: Zhao
authors.forename: Guang-hong
authors.initials: GH
authors.lastname: Cui
authors.forename: Min-tong
authors.initials: MT
authors.lastname: Xin
authors.forename: Shi-huan
authors.initials: SH
authors.lastname: Tang
mesh_terms: D000818:Animals; D017838:Bungarus / Q000145:classification / Q000235:genetics*; D045303:Cytochromes b / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004340:Drug Contamination; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D045303:Cytochromes b