Study on the identification of Testudinis Carapax et Plastrum Formula Granule by high-resolution melt-ing curve analysis
Objective To establish a high-resolution melting curve(HRM)identification for Testudinis Carapax et Plastrum(TCP)formula granule.Methods Based on the cytochrome C oxidase subunitⅠ(COI)sequence of Chinemys reevesii and its adulterant,specific primers were designed according to the specific single nucleotide polymorphisms(SNP)of Chinemys reevesii.The products were analyzed by HRM directly,the reaction was optimized and verified based on the optimal conditions.Results When the DNA template concentration was 0.62～374.88 ng/μL,the primer concentration was 0.1-0.4 μmol/L,the annealing temperature was 62℃and the cycle number was 45,the Tm value of TCP herbal decoction pieces and formula granule were(82.39±0.09)and(82.38±0.05)℃,respectively.No band was found in other adulterants and vinegar TCP formula granule.Conclusion The HRM method can quickly identify TCP crude medicine,herbal decoction pieces and formula granule,and also distinguish formula granule of TCP and its processed products,which provides a theoretical basis for the quality standard of Testudinis Carapax et Plastrum formula granule.
Testudinis Carapax et PlastrumFormula Granulehigh-resolution melting curvemolecular identification
万方数据
维普
