Specific PCR method for identification of Trionycis Carapax and its preparation.
10.19540/j.cnki.cjcmm.20181025.013
- Author:
Su-Qian CHENG
1
;
Yuan YUAN
1
;
Fu-Yan LIU
1
;
Chao JIANG
1
;
Yan JIN
1
;
Yu-Yang ZHAO
1
Author Information
1. State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Trionycis Carapax;
Trionycis Carapax Preparata;
molecular identification;
specific PCR
- MeSH:
Animals;
DNA Primers;
Polymerase Chain Reaction
- From:
China Journal of Chinese Materia Medica
2018;43(23):4569-4574
- CountryChina
- Language:Chinese
-
Abstract:
Trionycis Carapax is a commonly used animal medicine in Chinese medicine. It's difficult to identify Trionycis Carapax and its adulterants because of the loss of morphological characteristics after processing. To establish an efficient and stable method to identification Trionycis Carapax, this study combines SDS method with column purification to extract genomic DNA, uses universal primers for polymerase chain reaction (PCR) amplification and sequencing, and designs the specific primers based on the differences in the sequences of Pelodiscus sinensis and their adulterants. When the annealing temperature was 62 °C and the number of cycles was 35, the designed primer Biejia-272.F/R was used for PCR amplification and got optimum results. The crude drug and preparation of P. sinensis were all amplified to obtain a specific band of approximately 300 bp, while the adulterants showed no such a band. This method can be used as a rapid and accurate method to identify the authenticate of P. sinensis.
