| 大豆不同生长时期基因组DNA提取方法的优化 |
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| 引用本文： | 王月月,范龙,张洁,王冬梅.大豆不同生长时期基因组DNA提取方法的优化[J].中国农学通报,2013,29(36):319-325. |
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| 作者姓名： | 王月月 范龙 张洁 王冬梅 |
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| 作者单位： | 1. 河北农业大学生命科学学院2. |
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| 基金项目： | 转基因生物新品种培育重大专项子课题;河北省教育厅项目 |
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| 摘 要： | 以探索大豆嫩叶期基因组提取方法为基础，探索从富含酚类、多糖等代谢物质的大豆开花期叶片中获得高质量DNA的提取方法。采用不同的方法提取5种大豆品种在不同生长时期的叶片基因组，并通过对提取的DNA的含量测定、琼脂糖凝胶电泳分析和内参基因的PCR扩增，对不同方法所提取的DNA质量进行综合评价。在嫩叶期，利用前人改进的CTAB法能够分离出纯度高、质量好的DNA，而在开花期，采用本试验新改良的CTAB法可以更有效去除多糖、酚类等物质得到完整性和纯度较好的大豆基因组。前人改进的CTAB法和本试验摸索的新改良CTAB法分别适用于嫩叶期、开花期大豆基因组DNA提取，为大豆的分子生物学研究奠定了基础。
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| 关 键 词： | 大豆 不同生长时期 DNA提取方法 PCRi 多糖 |
| 收稿时间： | 2013/2/26 0:00:00 |
| 修稿时间： | 2013/4/18 0:00:00 |
Optimization of DNA extraction methods from soybean in different period |
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| Wang Yueyue,Fan Long,Zhang Jie,Wang Dongmei.Optimization of DNA extraction methods from soybean in different period[J].Chinese Agricultural Science Bulletin,2013,29(36):319-325. |
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| Authors: | Wang Yueyue Fan Long Zhang Jie Wang Dongmei |
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| Institution: | (College of Life Science, Agricultural University of Hebei, Baoding Hebei 071000) |
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| Abstract: | Using leaves in different growth periods of high-quality soybean varieties J-17, J-16, WX-1, WX-2 and Maverick as experimental materials, we used different methods for the genomic DNA extraction and determined the concentration of genome DNA. The DNA were tested by agarose gel electrophoresis, PCR amplification for internal control and comprehensive evaluation of the quality of different methods for DNA extraction. The results indicated that the DNA extracted with the modified CTAB method had higher quality and purity in fresh leaves period. The most of polysaccharides and polyphenolic compounds could be removed efficiently by utilizing new modified CTAB method and the new method was suitable for extracting high qualified DNA in flowering stage. This study laid the foundation of soybean moleculer bioresearch. |
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| Keywords: | Polysaccharides |
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