| 微量法提取高质量小麦DNA供大规模PCR分析 |
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| 引用本文： | 张志清,郑有良,王丕武,魏育明,颜泽洪,刘虹.微量法提取高质量小麦DNA供大规模PCR分析[J].西北农业学报,2005,14(6):83-86. |
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| 作者姓名： | 张志清 郑有良 王丕武 魏育明 颜泽洪 刘虹 |
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| 作者单位： | 1. 四川农业大学小麦研究所,四川,都江堰,611830
2. 吉林农业大学生物技术中心,吉林,长春,130118 |
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| 基金项目： | 四川省科技厅应用基础项目(2002A008)
四川省教育厅重点项目(03JY029-028-2863)
国家“863”项目(2003AA207100)资助 |
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| 摘 要： | 介绍了一种在提取小麦DNA实验中常用的CTAB提取方法基础上改良的微量DNA提取方法.通过提取多个材料的DNA并经分光光度计检测,琼脂糖电泳结果表明,这种方法提取的DNAOD260/280在2.00～1.8间波动,较常规大量法提取DNA的质量高,能满足100～200余次的PCR反应的需要.采用该法对提取的普通小麦DNA以及转抗除草剂基因Bar的转基因植株DNA分别进行SSR标记检测和PCR扩增,结果表明SSR标记扩增效果与大量法无明显的差异,抗性植株DNA能扩增出Bar基因的特征带.
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| 关 键 词： | 小麦 微量提取 DNA PCR分析 |
| 文章编号： | 1004-1389(2005)06-0083-04 |
| 收稿时间： | 2005/5/20 0:00:00 |
| 修稿时间： | 2005年5月20日 |
The Micromethod of Isolating High-quality Wheat Genomic DNA for Large-scale PCR Analysis |
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| ZHANG Zhi-qing,ZHENG You-liang,WANG Pi-wu,WEI Yu-ming,YAN Ze-hong and LIU Hong.The Micromethod of Isolating High-quality Wheat Genomic DNA for Large-scale PCR Analysis[J].Acta Agriculturae Boreali-occidentalis Sinica,2005,14(6):83-86. |
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| Authors: | ZHANG Zhi-qing ZHENG You-liang WANG Pi-wu WEI Yu-ming YAN Ze-hong and LIU Hong |
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| Institution: | Triliceae Research Institute, Sichuan Agricultural University, Dujiangyan Sichuan 611830, China;Triliceae Research Institute, Sichuan Agricultural University, Dujiangyan Sichuan 611830, China;Biotechnology Research Center, Jilin Agricultural University, Changchun Jilin 130118, China;Triliceae Research Institute, Sichuan Agricultural University, Dujiangyan Sichuan 611830, China;Triliceae Research Institute, Sichuan Agricultural University, Dujiangyan Sichuan 611830, China;Triliceae Research Institute, Sichuan Agricultural University, Dujiangyan Sichuan 611830, China |
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| Abstract: | This article introduced a modified micromethod of isolating wheat genomic DNA based on the traditional CTAB method. The agarose electrophoresis suggested that the higher-quality DNA obtained by this micromethod(OD260/280 = 2. 00- 1. 8), Meanwhile, about one hundred to two hundred PCR reactions could be met. Then, this method have been applied for isolating DNA of transgenic wheat and common wheat (non-transgenic) to amplified the given fragment of the Bar gene and SSR markers identification, respectively. The results indicated the 310 bp target fragment was amplified on the transgenic wheat, and the similarity SSR amplifying results was detected between the DNA isolated by the micromethod and traditional CTAB method. |
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| Keywords: | Wheat Micro-isolation DNA PCR Analysis |
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