6种链霉菌基因组DNA提取方法比较  

Comparison among Six Methods of Genome DNA Extraction from Streptomyces

作　　者：刘炳辉[1,2] 曹远银[1] 闫建芳[2] 齐小辉[2] 程浩[1,2] 刘秋[2] 

机构地区：[1]沈阳农业大学植物保护学院,辽宁沈阳110161 [2]大连民族学院生命科学学院,辽宁大连116600

出　　处：《河南农业科学》2008年第10期86-89,共4页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金(30671398);辽宁省自然科学基金(20062189);辽宁省教育厅项目(2004F079);大连市青年基金(2005J22JH040)

摘　　要：采用优化SDS、SDS、优化CTAB、CTAB、液氮研磨、微波法分别提取委内瑞拉链霉菌基因组DNA,比较6种方法的差异,选出最佳提取方法分别提取S.fradiae,S.venezuelae,S.ambofa-ciens,S.glaucescens,S.coelicolor基因组DNA,并利用16S rDNA的通用引物扩增相应的目的基因。结果表明:6种方法制备的基因组DNA以优化SDS法和微波法为上选,后者的纯度高但量很少。优化SDS法是6种提取方法中最可靠的DNA提取方法,DNA量大,纯度高,可靠,可作为PCR反应的模板进行16S rDNA基因有效扩增。Genomic DNA from S. f radiae , S. venezuelae , S. ambo f aciens , S. g laucescens , S. coelicolor was extracted by six methods including improved SDS method, SDS method, improved CTAB method,CTAB method, liquid nitrogen grinding method and microwave based method. The purity and output of the products were detected by PCR. The result showed that Genomic DNA from the six strains of actinomyces all could be obtained by the six methods, but the yield varied. There was no significant different between improved SDS method and microwave based method in terms of the purity, except output. The purity and output of DNA extracted by improved SDS method was obviously higher. Extraction of DNA from actinomyces by the improved SDS method is a common and trustworthy method and can be adopted in the PCR.

关 键 词：放线菌 基因组DNA提取 PCR 16S RDNA 

分 类 号：Q943.2[生物学—植物学]