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目的建立适合中药材DNA条形码分析的DNA提取方法。方法以6种富含次生代谢物质的药用顽拗植物为材料,对7种DNA提取方法进行比较。结果CTAB法3对中药材具有通用性,操作简便、快速,提取的DNA浓度和合格率比其他对照方法高,能满足DNA条形码PCR扩增的要求。该方法主要特点:(1)用3%的CTAB浓度代替常用的2%CTAB;(2)采用1%的PVP与0.2%β-巯基乙醇共同去除杂质和防止DNA降解;(3)采用10000r/min离心15min去除蛋白质等杂质。结论CTAB法3是适合中药材DNA条形码研究的DNA提取方法。
Objective To establish a DNA extraction method suitable for DNA barcode analysis of Chinese herbal medicines. Methods Six medicinal recalcitrant plants with secondary metabolites were used as materials to compare the seven DNA extraction methods. Results The CTAB method 3 has the advantages of commonality to Chinese herbal medicines, simple and rapid operation, high DNA concentration and qualified rate compared with other control methods, and can meet the requirement of PCR amplification of DNA barcodes. The main features of the method are as follows: (1) replacing common 2% CTAB with 3% CTAB concentration; (2) using 1% PVP together with 0.2% β-mercaptoethanol to remove impurities and prevent DNA degradation; (3) min centrifugation 15min to remove impurities such as protein. Conclusion CTAB method 3 is a DNA extraction method suitable for DNA barcoding study of Chinese herbal medicines.