DNA提取4种中药材方法的筛选

Screening of DNA extraction methods of four Chinese medicinal materials

目的筛选龙胆Gentiana scabra Bunge、锁阳Cynomorium songaricum Rupr、天花粉Trichosanthes kirilowii Maxim.、菟丝子Cuscuta chinensis Lam.的DNA提取方法。方法通过试剂盒法、SDS法、高盐低p H法、CTAB法以及改良PVP法,对4种中药材DNA进行提取。紫外分光光度、琼脂糖凝胶电泳和PCR扩增进行检测,SPSS20.0软件进行统计学分析。结果改良PVP法所得4种中药材DNA的得率均较高。SDS法和改良PVP法提取的DNA条带较清晰,高盐低p H法提取者呈弥散状态,试剂盒法和CTAB法提取者几乎看不到条带。试剂盒法和改良PVP法提取的DNA可完全扩增ITS2和psb A-trn H序列,而其他3种方法未能完全扩增。结论改良PVP法高效简单,最适合提取这4种中药材DNA。

AIM To screen the DNA extraction methods of Gentiana scabra Bunge, Cynomorium songaricum Rupr, Trichosanthes kirilowii Maxim. and Cuscuta chinensis Lam. METHODS The DNA of four Chinese medic- inal materials was extracted by kit, SDS, high-salt low-pI-I, CTAB and modified PVP methods. Ultraviolet spectro- photometry, agarose gel electrophoresis and PCR amplification were adopted in the detection, then the statistical a- nalysis was made by SPSS20. 0. RESULTS The DNA yields of four Chinese medicinal materials obtained by modified PVP method were relatively high. The DNA bands extracted by SDS and modified PVP methods were rela- tive clear, while those extracted by high-salt low-pH method showed diffuse status, and almost no bands were found in the DNA extracted by kit and CTAB methods. The DNA extracted by kit and modified PVP methods could com- pletely amplify ITS2 and psbA-trnH sequences. CONCLUSION Modified PVP method is effective and simple, which is the most suitable for extracting DNA of these four Chinese medicinal materials.