Improvement of Extraction Method of Animal-derived DNA from Colla Corii Asini and Identification of Donkey-Derived Components / 中国药学杂志

ABSTRACT

OBJECTIVE: To extract microamounts of animal-derived DNA from the products of Colla Corii Asini boiled at high temperature, establish and optimize a rapid identification method of donkey-derived components in Colla Corii Asini by polymerase chain reaction(PCR), and establish a new molecular biological method for assessing the quality of Colla Corii Asini. METHODS: The donkey-derived genomic DNA was extracted by DNA purification column instead of phenol, chloroform and other toxic organic solvents in SDS-PK method, and the SDS-PK method was optimized with the donkey-derived genomic DNA. RESULTS: The optimum sample size of Colla Corii Asini was 0.20 g. High quality genomic DNA of Colla Corii Asini could be obtained quickly after digestion in water bath for 1 h and then purified by DNA purification column. The purity ranged from 1.70 to 1.80, and the concentration of Colla Corii Asini could reach (187.8±0.56)ng•μL-1. PCR amplification, cloning, and sequencing were performed using specific primers, and the similarity to GenBank's registered Donkey species (MG931481.1) was 100%. CONCLUSION: This study provides animal-derived genomic DNA fragments from deep-processed Colla Corii Asini and Colla Corii Asini products within 90 min. The purity and concentration of extracted DNA can meet the requirements of molecular biological identification of Colla Corii Asini. The established PCR method can quickly identify the scorpion-derived components in Colla Corii Asini. The cloned donkey specific gene fragment can be used as a standard positive control to identify the authenticity of Colla Corii Asini. It is expected that it will be widely used in the quality supervision of Colla Corii Asini and related products.