Application of rapid PCR to authenticate medicinal snakes.
- Author:
Kang CHEN
;
Chao JIANG
;
Yuan YUAN
;
Lu-Qi HUANG
;
Man LI
- Publication Type:Journal Article
- MeSH:
Animals;
China;
DNA Primers;
genetics;
Polymerase Chain Reaction;
methods;
Reptilian Proteins;
genetics;
Snakes;
classification;
genetics
- From:
China Journal of Chinese Materia Medica
2014;39(19):3673-3677
- CountryChina
- Language:Chinese
-
Abstract:
To obtained an accurate, rapid and efficient method for authenticate medicinal snakes listed in Chinese Pharmacopoeia (Zaocysd humnades, Bungarus multicinctus, Agkistrodon acutus), a rapid PCR method for authenticate snakes and its adulterants was established based on the classic molecular authentication methods. DNA was extracted by alkaline lysis and the specific primers were amplified by two-steps PCR amplification method. The denatured and annealing temperature and cycle numbers were optimized. When 100 x SYBR Green I was added in the PCR product, strong green fluorescence was visualized under 365 nm UV whereas adulterants without. The whole process can complete in 30-45 minutes. The established method provides the technical support for authentication of the snakes on field.
