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| 高质量水稻基因组文库构建的策略和方法 | |
| 引用本文： | 梁卫红,刘一鸣,吴乃虎.高质量水稻基因组文库构建的策略和方法[J].河南师范大学学报(自然科学版),2006,34(2):177-179. |
| 作者姓名： | 梁卫红 刘一鸣 吴乃虎 |
| 作者单位： | 1. 河南师范大学,生命科学学院,河南,新乡,453007;中国科学院,遗传与发育生物学研究所,北京100080 2. 中国科学院,遗传与发育生物学研究所,北京100080;北京大学,生命科学学院,北京,100871 3. 中国科学院,遗传与发育生物学研究所,北京100080 |
| 基金项目： | 国家重点基础研究发展计划(973计划) , 高比容电子铝箔的研究开发与应用项目 , 河南省高校青年骨干教师资助项目 , 河南师范大学校科研和教改项目 |
| 摘 要： | 采用改良的CTAB法提取水稻基因组DNA,经Sau3AI部分酶切的产物经透析袋电洗脱法回收、正丁醇浓缩后,以1∶1的摩尔数比和载体臂连接.包装后测定,文库滴度达到106pfu/ml;随机酶切重组克隆显示,插入片段达到预期的15~23 kb之间,提示该方法适于植物材料高质量基因组文库的构建,有利于分离完整的基因序列及其调控区. |
| 关 键 词： | 基因组DNA 部分酶切 电洗脱 滴度 |
| 文章编号： | 1000-2367(2006)02-0177-03 |
| 收稿时间： | 2005-11-02 |
| 修稿时间： | 2005年11月2日 |
Strategy and Methods of Construction High Quality Rice Genomic Library |
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| LIANG Wei-hong,LIU Yi-ming,WU Nai-hu.Strategy and Methods of Construction High Quality Rice Genomic Library[J].Journal of Henan Normal University(Natural Science),2006,34(2):177-179. | |
| Authors: | LIANG Wei-hong LIU Yi-ming WU Nai-hu |
| Institution: | 1. College of Life Sciences, Henan Normal University, Xinxiang 453007,China;2 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China;3 College of Life Sciences,Peking University,Beijing 100871, China |
| Abstract: | Rice genomic DNA had been isolated by improved CTAB method,and the Sau3AI partial-digestion products were recovered by electric elution with dialysis bag.After concentrated by n-butyl alcohol,the purified partial-digestion products were ligated with vector arms in the 1∶1 ratios.After packaging,the titer reached to 10~6 pfu/ml.Quality detection showed that the DNA inserts were between 15~23 kb as expect,suggest that the method was appropriate for constructing plant genomic library,and was helpful to isolate full-length gene and its regulation sequences. |
| Keywords: | genomic DNA partial-digestion electric elution titer |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |