Fingerprint study of Perns et Testis Cervi mtDNA with PCR-SSCP
- Author:
Jing-Yu SUN
1
Author Information
- Publication Type:Journal Article
- Keywords: DNA; Penis et Testis Cervi; Polymerase chain reaction (PCR); Single strand conformation polymorphism (SSCP)
- From: Chinese Pharmaceutical Journal 2014;49(9):721-725
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To establish a method of molecular biology to identify the authenticity and varieties of Penis et Testis Cervi rapidly, simply and accurately. METHODS: The mitochondrial DNAs (mtDNA) of dried Cervus nippon Penis, Cervus elaphus. L Penis, Penis et Testis Bovis and commercial products were extracted and purified with column chromatography. A pair of primers were designed for PCR special amplification of mtDNA according to cytochrome b gene sequence in the cervidae mtDNA GenBank; the products of PCR were analyzed for the single strand conformation polymorphism (SSCP) by denaturing polyacrylamide gel electrophoresis (PAGE). RESULTS: The positive control samples and some commercial products showed clear single strand DNA (ssDNA) bands with different mobility, but the negative control sample and some other commercial products showed nothing. CONCLUSION: Column chromatography method is a good way for getting mtDNA with high purity and less destruction of structure for the follow-up special PCR amplification; SSCP technique not only shows ssDNA bands of mutually complementing clearly, but also reveals the difference of mobility among ssDNAs from all samples directly. For that reason, the technique of PCR-SSCP will be reliable and feasible to indentify the authenticity and varieties of products from cervidae.
