| 一种适宜拟南芥PCR检测的DNA提取方法（摘要）（英文） |
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| 引用本文： | 徐平丽,赵晋平,孟静静,李保龙,李新国,郭峰.一种适宜拟南芥PCR检测的DNA提取方法（摘要）（英文）[J].农业科学与技术,2010,11(3):41-42,155. |
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| 作者姓名： | 徐平丽 赵晋平 孟静静 李保龙 李新国 郭峰 |
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| 作者单位： | [1]山东省农业科学院高新技术研究中心,山东省作物与畜禽品种改良生物技术重点开放实验室,山东济南250100 [2]山东省曹县种子公司,山东菏泽274400 |
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| 基金项目： | 国家科技支撑计划，山东省优秀中青年科研奖励基金，山东省农业科学院青年基金 |
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| 摘 要： | 目的]介绍一种适于拟南芥PCR检测的DNA快速提取方法。方法]通过对常规DNA提取方法的改进（省去液氮研磨和苯酚提取的步骤）,获得可以快速大批量地提取拟南芥DNA样品的方法。于1.5ml Eppendorf管内加入400μl DNA提取液内含200 mmol/LTri（pH值7.5）,25 mmol/LEDTA（pH值8.0）,250 mmol/LNaCl,0.5% SDS（W/V）],剪取拟南芥叶片材料少许（1片以下）加入400μl DNA提取液,用微型研磨棒将叶片捣碎至溶液成绿色,放置3 ～5 min;加入400μl氯仿/异戊醇（体积比24：1）,混合均匀,12 000 r/min离心5 min;取上清液至另一1.5 ml Eppendorf管内,加入300μl异丙醇,颠倒混合均匀,室温下放置5 min,12 000 r/min离心5 min;弃去上清液,以70%乙醇漂洗,放置晾干,加入100μl灭菌超纯水溶解,4℃放置备用。以随机抽取的拟南芥转基因株系和突变株系为样品进行验证。结果]经过琼脂糖凝胶电泳及紫外吸收检测,DNA样品完整且污染少,PCR扩增目的片段结果良好,适于作为PCR反应的模板。经过对随机抽取的拟南芥转基因株系和突变株系的PCR检测,阳性植株目的基因扩增条带清晰,无假阳性,试验结果理想。结论]该方法适用于拟南芥DNA样品的快速提取、PCR检测及拟南芥突变体筛选工作。
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| 关 键 词： | 拟南芥 PCR DNA快速提取 |
A Rapid DNA Extraction Method for PCR Detection of Arabidopsis thaliana |
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| XU Ping-li,ZHAO Jin-ping,MENG Jing-jing,LI Bao-long,LI Xin-guo,GUO Feng.A Rapid DNA Extraction Method for PCR Detection of Arabidopsis thaliana[J].Agricultural Science ＆ Technology,2010,11(3):41-42,155. |
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| Authors: | XU Ping-li ZHAO Jin-ping MENG Jing-jing LI Bao-long LI Xin-guo GUO Feng |
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| Institution: | 1.Key Laboratory of Improved Biotechnology of Crops and Livestock and Poultry in Shandong Province,High-tech Research Center,Shandong Academy of Agricultural Sciences,Jinan 250100;2.Shandong Cao County Seed Company,Heze 274400 |
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| Abstract: | Objective] The aim was to introduce a rapid DNA extraction method for PCR detection of Arabidopsis thaliana.Method] Through the improvement of conventional DNA extraction method,a rapid Arabidopsis thaliana DNA extraction method was obtained.With randomly selected Arabidopsis thaliana transgenic strains and mutants as samples,the method was verified.Result] After electrophoresis,UV absorption detection,it was found that DNA samples are complete and less pollution,and the result of PCR amplification objective fragment was good which proved DNA is suitable as a template for PCR reaction.After PCR detection,positive plants gene amplified bands were clear,without false-positive,and the test results were satisfactory.Conclusion] The method is suitable for rapid extraction of Arabidopsis thaliana DNA and PCR detection. |
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| Keywords: | PCR Arabidopsis thaliana PCR DNA rapid extraction |
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