| 大肠杆菌质粒DNA提取方法的优化 |
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| 引用本文： | 王友如.大肠杆菌质粒DNA提取方法的优化[J].生物技术,2006,16(6):42-44. |
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| 作者姓名： | 王友如 |
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| 作者单位： | 湖北师范学院生物系,湖北,黄石,435002 |
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| 基金项目： | 国家自然基金面上项目资助(30570412) |
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| 摘 要： | 目的:简化操作流程,缩短提取时间,降低试验对操作者的危害。方法:该方法去除酚、氯仿等有害试剂,采用LiCl沉淀去除质粒制备物中小片段核酸(包括DNA和RNA);工程菌生长至对数期时通过加入氯霉素后不仅方便质粒DNA的提取过程中蛋白质的去除,而且可使质粒的拷贝数进一步增加,提高质粒DNA产量的目的。结果:实验改进方法所提取的质粒DNA产量高于常规方法,达到20μg/mL。结论:改进方法提取的质粒DNA,其下游的内切酶消化,PCR、重组质粒鉴定、转化大肠杆菌等实验的结果和重复性都令人满意,完全可用于一般的分子生物学研究。
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| 关 键 词： | 质粒 DNA提取 方法改进 纯化 |
| 文章编号： | 1004-311X（2006）06-0042-03 |
| 收稿时间： | 2006-07-28 |
| 修稿时间： | 2006-09-12 |
A Modified Method for Isolation of Plasmid DNA from E. coli |
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| WANG You-ru.A Modified Method for Isolation of Plasmid DNA from E. coli[J].Biotechnology,2006,16(6):42-44. |
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| Authors: | WANG You-ru |
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| Institution: | The Detartment of Biology, Hubei Normal University, Huangshi 435002, China |
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| Abstract: | Objective:To simplify the process of extraction of plasmid of DNA,shorten the time of extraction of plasmid of DNA,clear the damage to the manipulator.Methods: Hydroxybenzene and chloroform was excluded during the process,LiCl was used to precipitate the small fragments of DNA and RNA,chloramphenicol was applied to enhance the yield of plasmid DNA.Results:The yield of plasmid DNA obtained by the modified method was higher that obtained by normal one. Conclusion: The modified method is quite simple,rapid,economical and reliable.The quality of plasmid DNA prepared by this modified method can extensively meet the requirements for many molecuar biology experiments. |
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| Keywords: | plasmid extraction of DNA modified method purification |
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