| 优化的密度梯度离心法提取质粒 DNA |
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| 引用本文： | 高英杰,赵红桃.优化的密度梯度离心法提取质粒 DNA[J].衡水学院学报,2014(4):49-51. |
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| 作者姓名： | 高英杰 赵红桃 |
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| 作者单位： | 河北师范大学生命科学学院,河北石家庄050024 |
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| 摘 要： | 质粒常被用做基因的载体．纯化质粒DNA的方法可以分为3种：碱裂解法、煮沸裂解法和SDS碱裂解法．这些方法可以纯化共价闭合环结构质粒DNA，而大量获得高纯度的闭环结构质粒DNA最经典方法是氯化铯．溴化乙锭密度梯度离心法．该方法经过我们的实践与优化，减少了质粒中蛋白质和基因组DNA的污染，提高了质粒的纯度和得率，所得到的质粒用于拟南芥原生质体转化，效率高达60％～70％，明显高于普通的质粒小量抽提试剂盒所提取的质粒DNA．
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| 关 键 词： | 质粒DNA 氯化铯-溴化乙锭 密度梯度离心法 拟南芥原生质体 转化效率 |
Optimize the Extraction of Plasmid DNA with Method of Density Gradient Centrifugation |
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| GAO Ying-jie,ZHAO Hong-tao.Optimize the Extraction of Plasmid DNA with Method of Density Gradient Centrifugation[J].Journal of Hengshui University,2014(4):49-51. |
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| Authors: | GAO Ying-jie ZHAO Hong-tao |
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| Institution: | (College of Life Science, Hebei Normal University, Shijiazhuang, Hebei 050024, China) |
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| Abstract: | Plasmid is often used as gene carrier. Method of purifying plasmid DNA can be divided into three types： the alkaline lysis method, boiling lysis and SDS alkaline lysis method. These methods can be used to purify covalently closed circular plasmid DNA, and the classical method of obtaining high-purity closed circular plasmid DNA is cesium chloride - ethidium bromide density gradient centrifugation. Through practice and optimization, we can reduce the pollution of protein and genomic DNA in plasmid and raise the purity and yield. The obtained plasmid is used for the conversion of arabidopsis thaliana protoplasts, and the conversion efficiency is as high as 60%-70%, which is significantly higher than that extracted with common plasmid extraction test kit. |
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| Keywords: | plasmid DNA cesium chloride-ethidium bromide density gradient centrifugation arabidopsis thaliana protoplast conversion efficiency |
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