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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

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【作者】 孙慧慧董晓璇唐蕾刘海丽张峰杨瑞金

【Author】 SUN Hui-hui et al(Key Laboratory of Industrial Biotechnology of Ministry of Education,Jiangnan University,Wuxi,Jiangsu 214122)

【机构】 江南大学工业生物技术教育部重点实验室

【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。

【Abstract】 [Objective] The different extraction methods of Arthrobacter DNA were compared to obtain high-quality genomic 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 electrophoresis of the purified DNA extracted with modified CTAB showed that the genomic 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,economical 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
【Key words】 ArthrobacterDNA extractionPCR
【基金】 国家863计划项目(2006AA10Z336)
  • 【文献出处】 安徽农业科学 ,Journal of Anhui Agricultural Sciences , 编辑部邮箱 ,2009年27期
  • 【分类号】Q78
  • 【被引频次】2
  • 【下载频次】219
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