上海牡丹根际土壤DNA提取方法研究

Study on DNA Extraction Method in Rhizosphere Soil of Poeny in Shanghai

采用玻璃珠-溶菌酶法、蛋白酶K-SDS-热处理法、PVPP-溶菌酶-SDS法等3种土壤DNA提取方法,分别提取上海地区牡丹根际土壤DNA。玻璃珠-溶菌酶法、蛋白酶K-SDS-热处理法提取的DNA,含有大量的杂质,抑制PCR扩增反应,但分别用DNA溶液过柱纯化后不同倍数稀释和凝胶电泳回收纯化,都能扩增出相应的目的条带。PVPP-溶菌酶-SDS法提取的DNA杂质较少,直接用于PCR扩增反应,可以扩增出与预期一致的条带。PVPP-溶菌酶-SDS法不需要纯化,时间较短,可以有效地扩增出预期条带,适合上海地区牡丹根际土壤DNA的提取。

Three DNA extraction methods of glass bean-lysozyme method,proteinase K-SDS-heating method,PVPP-lysozyme-SDS method were used to extract DNA in rhizosphere soil of poeny in Shanghai.The soil DNA extracted by glass bean-lysozyme method and proteinase K-SDS-heating method included many impurities,which inhibited PCR amplification.After purified by agarose method,the soil DNA was used for PCR amplification.DNA extracted by method of PVPP-lysozyme-SDS included less impurities, which could be used for PCR amplification,not purifying.PVPP-lysozyme-SDS method for DNA extraction was simple and effective, which was suitable for rhizosphere soil of poeny in Shanghai.