DNA Extraction Method Optimization for Animal Medicines and Identification of Commercial Animal Medicines Using DNA Barcoding
- VernacularTitle:动物药材DNA提取方法优化和市售药材DNA条形码鉴定研究
- Author:
Xuzhao LIU
1
,
2
;
Jinxin LIU
;
Wei SUN
;
Jingyuan SONG
;
Linchun SHI
;
Zhiying SUN
Author Information
- Keywords: Animal medicines; DNA barcoding; DNA extraction; lysis buffer; EDTA
- From: World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(4):602-609
- CountryChina
- Language:Chinese
- Abstract: The DNA extraction method of animal medicine material is difficult and un-unified,which limits the application of molecular identification to identify animal medicines.In this study,based on the DNA extraction theory of SDS,we assessed the effects of three elements including different EDTA concentrations (0.025 mol·L-1,0.25 mol· L-1,and 0.5 mol· L-1) and whether containing NaCl and Triton X-100 in the lysis buffer on the quality of DNA extracted from different kinds of animal medicine.The optimized lysis buffer was used to extract DNA from 121 commercial animal medicines for original and species identification.The results showed that the lysis buffer of 1％ SDS,0.03 mol· L-1 Tris-HCl,0.25 mol· L-1 EDTA and 0.2 mol· L-1 NaCl had the optimum effect on DNA extraction.This lysis buffer can obtain DNA from animal medicine which is difficult to extract,such as Cicadae periostracum.The DNA extractions of 121 commercial animal medicines by optimized lysis buffer can satisfy the experimental requirements for molecular identification.All samples of commercial animal medicines can be accurately identified to the level of species.It was concluded that optimized lysis buffer can be used in the DNA extraction of different kinds of animal medicines except shells,secretions and processed products.This method provides technique support for the molecular identification of animal medicines.
