中国中药杂志

2014, v.39(19) 3678-3683

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碱裂解法快速提取中药炒制品DNA的研究
Rapid extraction of DNA from Chinese medicinal products by alkaline lysis

郑琪;蒋超;黄璐琦;张志杰;李娆娆;陈康;袁媛;金艳;
ZHENG Qi;JIANG Chao;HUANG Lu-qi;ZHANG Zhi-jie;LI Rao-rao;CHEN Kang;YUAN Yuan;JIN Yan;Shaanxi University of Chinese Medicine;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;Institute of Chinese Materia Medica,Chinese Academy of Chinese Medical Sciences;Anhui College of Traditional Chinese Medicine;

摘要(Abstract):

该文旨在探索一种适用于中药炒制品DNA快速提取的方法。以氢氧化钠,1%PVP40和1%Triton X-100配制成碱裂解缓冲液,Tris-HCl为中和液,经加热裂解和中和2步提取不同炒制方法制备的中药炮制品的DNA,选择2种方法对DNA进行纯化,并以纯化后DNA作为模板利用通用引物进行PCR扩增。结果表明优化碱裂解法可简单快速的提取出药材DNA,槐米炒制品DNA质量浓度为(420.61±123.91)g·L-1,且使用5%Chelex-100树脂纯化可以提高DNA提取浓度。研究结果证明优化碱裂解法适用于中药炒制品的DNA快速提取。
The study is aimed to explore a rapid method to extract DNA from fried Chinese medicinal products. The alkaline lysis buffer was made of sodium hydroxide,1% PVP and 1% Triton X-100 and Tris-HCl solution was neutralized,through heat cracking and neutralization two step to extract DNA from processed and prepared products of traditional Chinese medicine. Then universal primes were used to amplify PCR products for fired Chinese medicinal materials. The results indicated the optimized alkaline lysis method for extracting DNA is quick and easy. Extracting of the different processed Sophora japonica of DNA concentration was( 420. 61 ± 123. 91)g·L- 1. Using 5% Chelex-100 resin purification can improve the DNA concentration. Our results showed that the optimized alkaline lysis method is suitable for Chinese medicinal materials for quickly DNA extraction.

关键词(KeyWords): 中药炒制品;DNA快速提取;碱裂解法;Chelex-100
Chinese medicinal fried products;quickly DNA extraction;alkalinelysis;Chelex-100

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基金项目(Foundation): 中医药行业研究专项(201407003)

作者(Author): 郑琪;蒋超;黄璐琦;张志杰;李娆娆;陈康;袁媛;金艳;
ZHENG Qi;JIANG Chao;HUANG Lu-qi;ZHANG Zhi-jie;LI Rao-rao;CHEN Kang;YUAN Yuan;JIN Yan;Shaanxi University of Chinese Medicine;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;Institute of Chinese Materia Medica,Chinese Academy of Chinese Medical Sciences;Anhui College of Traditional Chinese Medicine;

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