适用于竹林土壤PCR-DGGE分析用的微生物总DNA提取及纯化方法

为了获得完整、高纯度的竹林土壤微生物总DNA，采用改良的十二烷基硫酸钠-高盐抽提法对5种竹林土壤进行DNA提取，通过DNA凝胶回收试剂盒纯化粗提的DNA，利用细菌16 SrDNA基因的V3区引物对纯化的DNA进行了聚合酶链式反应一变性梯度凝胶电泳（PCR-DGGE）验证。结果表明，该方法所需土壤样品少，抽提的DNA片段均在23kb以上，完整性好；采用DNA凝胶回收试剂盒纯化能够有效去除粗提DNA中大部分杂质，获得高质量的DNA；以此DNA为模板进行DGGE检测所反映的微生物信息量丰富。图3表1参15

Extraction and purification of microbacterial total DNA from bamboo soil for PCR-DGGE analysis

An improved SDS-extraction method was used to obtain high quality DNA from five different bamboo soil samples. The crude DNA was purified by a simple method with DNA gel extraction kit, amplified with 16 S rDNA-based primers in V3 region by use of the polymerase chain reaction （PCR）, and then the products of PCR was identified by denature gradient gel electrophoresis （DGGE）. The results showed that the DNA extracted from a little soil by this method was greater than 23 kb in size. The simple purification method we described here can remove greater part of contaminants from crude DNA and obtain high purity DNA. DGGE analysis patterns of amplified DNA presented abundant fragments. The extraction and purification method was considered to be an effective method for DNA isolation and purification from bamboo soil. It can be used to help characterize the biological composition and diversity of the microbial population in bamboo soil samples. Ch, 3 fig. 1 tab. 15 ref.]