节杆菌基因组DNA提取方法的改良  

Improvement of the Extraction Method of the Genome DNA from Arthrobacter

作　　者：孙慧慧[1] 董晓璇[1] 唐蕾[1] 刘海丽[1] 张峰[1] 杨瑞金[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122

出　　处：《安徽农业科学》2009年第27期12934-12935,共2页Journal of Anhui Agricultural Sciences

基　　金：国家863计划项目(2006AA10Z336)

摘　　要：[目的]研究比较了节杆菌的不同DNA提取方法,以获得高质量的基因组DNA。[方法]采用反复冷冻研磨、甲苯处理对细胞进行预处理,随后进行CTAB抽提,并与细菌基因组提取试剂盒进行比较。[结果]改良的CTAB法提取的DNA,纯化之后经电泳检测表明,基因组DNA条带清晰,量度均一,DNA片断大小一致;以上述DNA为模板进行PCR扩增,条带单一、特异性强,其中利用经甲苯方法提取的DNA快速、简便、经济实用,可作为PCR扩增的模板进行后续的基因操作和分子生物学研究。[结论]改良的CTAB法能够有效地提取节杆菌基因组DNA,纯度达到要求。[ Objective ] The different extraction methods of Arthrobacter DNA were compared to obtain high-quality genomie DNA. [ Method ] The cells were pretreated in repeated freeze-grinding and toluene; then the DNA was extracted with CTAB ion, and finally, the result was compared with the bacterial genomic comparison kit. [ Results] The result of the agarose gel eleetrophoresis of the purified DNA extracted with modified CTAB showed that the genomie DNA bands were clear, homogeneity in measure and same size of DNA fragment. The DNA as a template was amplified by PCR, it had single DNA band and specific trait, in which, the extraction method of DNA pretreated by toluene was rapid, simple, eeono＇mieal and practical, which could be used as a template for PCR amplification and for the follow-up genetic manipulation and molecular biology research. [ Conclusion] The modified CTAB method was effectively in the extraction of Arthrobacter genome DNA, which purity was required for follow-up research.

关 键 词：节杆菌 DNA提取 PCR 

分 类 号：Q55[生物学—生物化学]