高山被孢霉DNA提取方法的比较与改进  

Comparison and Improvement of the Methods of DNA Extraction from Mortierella alpina

作　　者：程国旺[1] 吴中华[2] 徐剑[2] 谷运红[1] 余增亮[1] 

机构地区：[1]中国科学院离子束生物工程学重点实验室等离子体物理研究所,合肥230031 [2]合肥丰乐种业股份有限公司,合肥230031

出　　处：《安徽农业大学学报》2004年第2期203-206,共4页Journal of Anhui Agricultural University

基　　金：国家自然科学基金(119890300);安徽省自然科学基金项目(99047640)资助。

摘　　要：利用CTAB法、SDS裂解法、氯化苄法和溶菌酶消化法提取高山被孢霉菌团的DNA。结果表明,CTAB法提取的DNA收率与纯度均较高,收率在700μg·g-1菌团以上,蛋白质污染少;SDS裂解法提取的DNA收率和纯度均较低,蛋白质污染严重;氯化苄法提取的DNA纯度较高,且无需苯酚纯化,但其易降解成DNA片段,质量稳定性较差;溶菌酶消化法无需采用液氮研磨,提取的DNA能达到分子生物学分析的要求,但收率较低,且步骤烦琐,成本较高。用改进的CTAB法可得到高纯度、高收率的被孢霉DNA,并可用于酶切或PCR扩增。The effect of 4 methods of DNA extraction from Mortierella alpina was compared.The results of the experiments showed that a higher yield and better quality of DNA could be obtained by CTAB method (the lysis agent was cetyltrimethyl ammonium bromide) ,which got to more than 700 μg·g^(-1) fresh mycelia mass with little protein impurities. On the contrary, a lower yield and quality of DNA was extracted by the SDS method (the lysis agent was sodium dodecyl sulfate), which contained a lot of protein impurities. Benzyl chloride was not only a kind of cell lysis agent, but also it could purify the extracted total DNA.Thus, a high-quality DNA could be produced without the presence of phenol under this protocol. However, it tended to cause lower yield and more DNA digestion. The lysozyme digestion was another technique of DNA extraction. The DNA extracted by this protocol was suitable for the usual molecular biological experiments, but it only produced a lower yield of DNA and higher cost. High quality and quantity of DNA from the fungi could be provided by the modified procedure with CTAB, which was practical for isolating DNA from a large number of fungal samples. The extracted DNA was suitable for RAPD-PCR and EcoRⅠdigestion analysis.

关 键 词：DNA提取 高山被孢霉 真菌 

分 类 号：Q949.320.6[生物学—植物学]