蒙药材阿给(小白蒿)基因组DNA的提取

Genomic DNA Extraction of Mongolian Medicine Artemisia Frigida Willd.

为获得能用于蒙药材阿给(小白蒿)RAPD分析及其他分子生物学研究的高质量DNA,选用经典的CTAB法、改进的CTAB法和高盐低pH法提取DNA,研究和改进几种DNA提取法。由于蒙药材含有大量的多糖和酚类物质,改进法对样品进行了预处理:先将样品与PVP和VitC共研碎,再用Tris-HCl缓冲液洗涤样品,离心的沉淀用于进行下一步操作。通过电泳、紫外吸收A260nm/A280nm和PCR扩增检测所得DNA的产量和质量。结果表明,改进的CTAB法和高盐低pH法都能获得高质量的DNA进行PCR扩增,其中以改进的CTAB法效果最好。

To extract high quality DNA for RAPD and other molecular biology research from Mongolian medicine Agi （Artemisia Frigida Willd. ） ,classic CTAB method, improved CTAB method and low pH medium with high salt （LPHS） method were employed for DNA extraction. Because of the Mongolian medicine containing a large number Of polyphenols and polysaccharides, improved methods add pretreatment step as the following： samples were ground with PVP and VitC by using liquid nitrogen, then washed by Tris-HCl buffer, centrifugal sedimentation used for the next operation. The yield and quality of DNA was analyzed by agarose gel electrophoresis, ratios of A260nm/A280nm and PCR amplification. The results were showed that improved CTAB method and LPHS method can produce high quality DNA for PCR amplification. Improved CTAB method is the best of the three.