七叶树叶片的DNA提取

Extraction of DNA from Leaves of Aesculus chinensis

[目的]探索能用于RAPD分析的七叶树叶片DNA的提取方法。[方法]以七叶树嫩叶为材料,使用聚乙烯吡咯啉酮(PVP)和巯基乙醇方法从七叶树叶片中提取DNA。[结果]聚乙烯吡咯啉酮被用于去除多酚,巯基乙醇是强烈的还原剂和蛋白质变性剂,提取过程中聚乙烯吡咯啉酮和巯基乙醇的使用促进了DNA质量和提取效率的提高。遵循这种程序提取的DNA需通过检查,质量好的方可用于RAPD分析,PCR扩增次数可达500次左右。应该根据实际情况调整聚乙烯吡咯啉酮和巯基乙醇的用量。[结论]使用聚乙烯吡咯啉酮和巯基乙醇方法得到的七叶树叶片DNA完全能满足随机扩增多态性DNA分析的要求。

[ Objective] The aim was to explore the extraction method of DNA used for RAPD analysis from leaves of Aesculus chinensis. [ Method ] With the tender leaves of A. chinensis as materials, the DNA was extracted from the leaves of A. chinensis by the method of polyvi- nyl pyrrolidone （PVP） and mercaptoethanol. [ Result] PVP was used to eliminate the polyphenol and the mereaptoethanol was the strong reducing agent and protein denaturant. Using PVP and mercaptoethanol during the extraction process improved the DNA quality and the extraction efficiency. According to the procedure the extracted DNA needed to pass the detection, the DNA with good quality could be used for RAPD analysis and the times of PCR amplification could reach about 500. The doge of PVP and mercaptoethanol should be regulated according to the actual condition. [ Conclusion ] The DNA from the leaves of A. chirtertsis extracted by the method of PVP and mercaptoethanol could meet the demand of the analysis of the random amplified polymorohie DNA.