| 碱裂解法提取P.ananatis变异体质粒DNA改良研究 |
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| 引用本文： | 徐艳,李茵. 碱裂解法提取P.ananatis变异体质粒DNA改良研究[J]. 安徽农业科学, 2011, 39(16): 9522-9524 |
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| 作者姓名： | 徐艳 李茵 |
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| 作者单位： | 东华大学环境科学与工程学院,上海,201620;东华大学环境科学与工程学院,上海,201620 |
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| 基金项目： | 上海市重点学科建设项目(B604) |
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| 摘 要： | [目的]对常规碱裂解法进行改进。[方法]分别改变溶液Ⅰ、Ⅱ、Ⅲ及无水乙醇的反应时间,少量提取质粒DNA。根据产率和纯度,得出各溶液的最佳反应时间,以紫外分光光度法、琼脂糖凝胶电泳结果验证提取效果。[结果]将常规碱裂法中溶液Ⅰ、Ⅱ、Ⅲ及无水乙醇的反应时间由原来的5、5、5、15 min分别缩短至0、1、1、5 min,纯度达1.8~1.9,产率达180.0~248.0μg/ml。[结论]改良法提取质粒DNA,总提取时间缩短了23 min。
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| 关 键 词： | 质粒DNA 改良碱裂解法 提取时间 |
Improvement of Alkaline Lysis Method on Extracting Plasmid DNA of Transposon Mutants in P.ananatis |
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| XU Yan et al. Improvement of Alkaline Lysis Method on Extracting Plasmid DNA of Transposon Mutants in P.ananatis[J]. Journal of Anhui Agricultural Sciences, 2011, 39(16): 9522-9524 |
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| Authors: | XU Yan et al |
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| Affiliation: | XU Yan et al(College of Environmental Science & Engineering,Donghua University,Shanghai 201620) |
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| Abstract: | [Objective] The alkaline lysis method was improved.[Method] To extract a small amount of plasmid DNA,the reaction time of solution Ⅰ,Ⅱ,Ⅲ and ethanol was changed,based on the yield and purity,the optimum reaction time was obtained.The purification efficiency in the experiment was to be tested by UV spectrophotometer examination and agarose gel electrophoresis.[Result] Decrease the reaction time of solution Ⅰ,Ⅱ,Ⅲ and ethanol from 5,5,5,15 min to 0,1,1,5 min respectively,the purity was between 1.8 and 1.9,the yield was between 180.00 and 248.0 μg/ml.[Conclusion] The overall DNA extraction time using improved alkaline lysis method was reduced by 23 minutes. |
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| Keywords: | Plasmid DNA Modified alkaline lysis method Extraction time |
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