| 基于磁珠的细菌基因组DNA快速提取方法 |
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| 引用本文： | 高秋月,景奉香,李海燕,陈季武,金庆辉,赵建龙.基于磁珠的细菌基因组DNA快速提取方法[J].安徽农业科学,2010,38(21):11071-11074. |
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| 作者姓名： | 高秋月 景奉香 李海燕 陈季武 金庆辉 赵建龙 |
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| 作者单位： | 华东师范大学生命科学学院,上海,200062;中国科学院上海微系统与信息技术研究所,上海,200050;中国科学院上海微系统与信息技术研究所,上海,200050;华东师范大学生命科学学院,上海,200062 |
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| 基金项目： | 中国科学院知识创新工程重要方向项目 |
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| 摘 要： | 目的]为临床上大肠杆菌O157∶H7的快速检测提供参考。方法]依据高盐离子浓度下多聚核苷酸与羧基修饰的磁珠表面可发生非特异性吸附,而蛋白、单糖、多糖和脂类等细胞成分不被吸附的原理,建立了一种以磁性微珠为载体的细菌基因组DNA纯化方法—磁性DNA纯化法。结果]该方法可在7min内完成对大肠杆菌O157∶H7基因组DNA的高效富集与纯化;与德国进口同类试剂盒gene-MAG-DNA/Bacteria及AxyPrep细菌基因组DNA纯化试剂盒相比,该方法操作简单、耗时短、不需要使用离心机和有机试剂,且无需常规快速提取法的沸水浴等步骤,安全、快速、成本低,在临床、食品、环境病原体快速检测领域具有巨大的推广价值。结论]该研究成功建立了一种细菌基因组DNA的快速提取方法。
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| 关 键 词： | 磁珠 DNA纯化 大肠杆菌O157:H7 |
Rapid Extraction Methods of Bacterial Genome DNA Based on Magnetic Bead |
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| Institution: | GAO Qiu-yue et al(School of Life Science,East China Normal University,Shanghai 200062) |
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| Abstract: | Objective] The research aimed to provide reference for clinical rapid detection of Escherichia coli O157∶H7.Method] Based on the principle that the nonspecific adsorption could occur on the surface of magnetic bead modified by polynucleotide and carboxy group under high salt concn.and the cell components including protein,monosaccharide,polysaccharide and lipoid could not be adsorbed,the magnetic DNA purification-a purification method of bacterial genome DNA was established with magnetic superbead as vector.Result] The efficient enrichment and purification of genome DNA from E.coli O157∶H7 could be finished in 7 min by using this method.Compared with the same kind of kit geneMAG-DNA/Bacteria and purification kit of AxyPrep bacterial genome DNA imported from Germany,this method was easy and convenient to operate,time-saved,needed no centrifugal machine and organic solvents and the steps such as boiling-water bath of conventional rapid extraction methods were unnecessary.This method was safe,rapid and low-cost,so it had enormous generalization value in rapid detection domain of pathogens in clinic,food and environment.Conclusion] A rapid extraction method of bacterial genome DNA was successfully established in this research. |
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| Keywords: | Magnetic bead DNA purification Escherichia coli O157∶H7 |
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