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| 梅花基因组AFLP银染反应体系的建立和优化 | |
| 引用本文： | 明军,张启翔,晏小兰,邱丽娟.梅花基因组AFLP银染反应体系的建立和优化[J].北京林业大学学报,2003,25(3):17-21. |
| 作者姓名： | 明军 张启翔 晏小兰 邱丽娟 |
| 作者单位： | 1. 湖北农学院园艺系 2. 北京林业大学园林学院 3. 中国梅花中心,武汉 4. 中国农科院品种资源所 |
| 基金项目： | 湖北省自然科学基金;2001ABB118; |
| 摘 要： | 采用改进的SDS DNA提取方法从梅花的嫩叶和老叶中提取获得总DNA .所得DNA样品的OD2 6 0 OD2 80 值在1 7～ 1 9之间 ,琼脂糖凝胶上主带清晰、较少降解、样品纯度高、蛋白去除干净且得率高 .该体系对原提取程序作了多处简化 ,使操作简便、快捷、高效 ,适合于梅花DNA提取 ,所提取的DNA样品不含PCR反应抑制剂及其他酶反应抑制剂 ,可被限制性内切酶MseⅠ和EcoRⅠ完全酶切 ,适合AFLP PCR反应应用 ,得到清晰的指纹式样 . |
| 关 键 词： | 梅花 DNA提取 AFLP DNA模板 银染 |
DNA template preparation and technology system building for AFLP analysis of Prunus mume |
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| Ming Jun,Zhang Qixiang,Yan Xiaolan,Qiu Lijuan.DNA template preparation and technology system building for AFLP analysis of Prunus mume[J].Journal of Beijing Forestry University,2003,25(3):17-21. | |
| Authors: | Ming Jun Zhang Qixiang Yan Xiaolan Qiu Lijuan |
| Institution: | Ming Jun,Zhang Qixiang,Yan Xiaolan,Qiu Lijuan. Department of Horticulture,Hubei Agricultural College,Jingzhou,434025,P. R. China. |
| Abstract: | The improved method of SDS DNA isolation is used to extract total DNA from tender or old leaves of \%Prunus mume\% in this study. High quality total DNA from plant was isolated in less than 3 hours. The values of OD 260 / OD 280 of each sample were between 1 7 and 1 9 Strips in agarose electrophoresis were clear and less decomposed. DNA samples with high purification and high yield rate could be gained by this method. It is much more simplified than former methods at many steps. The procedure of the method is easy, fast and effective, and is good to extract genomic DNA of \%Prunus mume\%. These genomic DNA samples are intact without contaminating nucleases or inhibitor. The DNA isolated with this method could be completely digested by restriction enzyme of Mse Ⅰ and EcoR Ⅰ, and good enough for use in following applications including PCR and analysis of AFLP. The systems suitable for silver stained AFLP analysis were established and capable of producing sharp fingerprint patterns. |
| Keywords: | Prunus mume DNA isolation AFLP DNA template silver staining |
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