| 质粒DNA小量提取法的改进 |
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| 引用本文： | 姚伟,周会,徐景升,余爱丽,张木清,陈如凯.质粒DNA小量提取法的改进[J].应用与环境生物学报,2005,11(6):776-778. |
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| 作者姓名： | 姚伟 周会 徐景升 余爱丽 张木清 陈如凯 |
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| 作者单位： | 1. 三峡大学生物技术研究中心,湖北,宜昌,443002
2. 福建农林大学农业部甘蔗生理生态与遗传改良重点开放实验室,福州,350002 |
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| 基金项目： | 国家"863"计划项目(2002AA241031),国家自然科学基金(No.30170590)资助~~ |
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| 摘 要： | 介绍了一种改进的小量提取质粒DNA的方法,该方法用NH4Ac和LiCl代替常规方法中苯酚和氯仿的抽提,有效去除了样品中RNA和蛋白质;用95%乙醇代替无水乙醇沉淀质粒DNA,降低了终产物中盐和多糖等杂质的含量,获得的质粒DNA质量好,产率高,能满足常规分子生物学实验的要求,如PCR、酶切、转化大肠杆菌以及植物遗传转化.图4表1参7
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| 关 键 词： | 质粒DNA 碱裂解法 提取方法 |
| 收稿时间： | 2004-10-18 |
| 修稿时间： | 2004年10月18 |
AN IMPROVED METHOD FOR MINIPREP OF PLASMID DNA |
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| YAO Wei,ZHOU Hui,XU Jingsheng,YU Aili,ZHANG Muqing,CHEN Rukai.AN IMPROVED METHOD FOR MINIPREP OF PLASMID DNA[J].Chinese Journal of Applied and Environmental Biology,2005,11(6):776-778. |
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| Authors: | YAO Wei ZHOU Hui XU Jingsheng YU Aili ZHANG Muqing CHEN Rukai |
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| Abstract: | An improved protocol for isolation of plasmid DNA from E. coli was described. The improved method used NH_ 4Ac and LiCl instead of phenol and chloroform extraction, and used 95% alcohol instead of absolute alcohol, which could wipe off large amounts of RNA, protein, salt, amylase and other undesired materials. The experiments demonstrated that the improved method generated good quality plasmid DNA with high yield, and it could meet the standards for many common molecular biology experiments, such as PCR, restriction enzyme digestion, E. coli transformation and plant genetic transformation. Fig 4, Tab 1, Ref 7 |
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| Keywords: | plasmid extraction alkaline lyses method for extraction |
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