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机构地区:[1]中国高校工业微生物资源和信息中心及江南大学工业生物技术教育部重点实验室
出 处:《食品与发酵工业》2007年第5期29-31,共3页Food and Fermentation Industries
基 金:国家自然资源科技平台(2005DKA21208);教育部科技基础条件平台(505006)
摘 要:16S rDNA基因同源性分析是一种常用的细菌分子鉴定方法,16S rDNA基因的扩增需要细菌染色体DNA作为模板。传统的细菌染色体DNA提取方法费时费力,限制了细菌分子鉴定的规模化。文中建立了1种新的快速高效的细菌染色体DNA提取方法,整个提取过程仅耗时20min,从而使得细菌的快速鉴定成为可能。用该方法制备的染色体DNA无需任何处理即可作为模板用于PCR扩增细菌的16S rDNA基因,并获得了良好的扩增结果以及扩增产物的测序结果。16S rDNA sequence-based alignment and analysis is a well-received and well-applied method tor bacterial identification and classification. To carry out technigue efficiently, the method should include a well-developed, rapid and repeatable protocol for extraction and preparation of target bacterial chromosomal DNA in order to serve as template for amplification of 16S rDNA by PCR techniques. However, the present methods for the bacterial chromosomal DNA extraction are inefficient in time and insufficient for large amount of samples. In the present article, a novel rapid method for the bacterial chromosomal DNA preparation has been developed and successfully applied for rapid identification of a large amount of bacterial isolates. It only spent 20 minutes for preparation of bacterial chromosomal DNA and the extracted chromosomal DNAs were directly used as template for amplification of 16S rDNA. Subsequent PCR products were of quantity for sequencing analysis.
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