| 大白菜基因组DNA的提取及AFLP反应体系的建立(英文) |
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| 引用本文： | 孟淑春,张海英,郑晓鹰,刘玉梅,王永健.大白菜基因组DNA的提取及AFLP反应体系的建立(英文)[J].Agricultural Science&Technology,2009(3). |
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| 作者姓名： | 孟淑春 张海英 郑晓鹰 刘玉梅 王永健 |
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| 作者单位： | 国家蔬菜系统工程技术研究中心;中国农业科学院蔬菜花卉研究所; |
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| 基金项目： | Supported by Construction and Industrialization Development of Beijing Vegetable Germplasm Modified Center(H022020130130);;the Great Project of Natural Science Foundation of Beijing (5050001)~~ |
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| 摘 要： | Objective] The obtained clear AFLP fingerprint of Chinese cabbage provided basis for studies on the molecular markers of Chinese cabbage cultivars and the phylogenetic relationship among Chinese cabbage cultivars. Method] With the test materials of leaves of Chinese cabbages, the high-quality total DNA from leaves of Chinese cabbages was extracted by the modified CTAB method. DNA restriction-ligase reaction, pre-amplification and selective amplification were optimized, and the AFLP silver-staining reaction system for Chinese cabbage was established. Result] The quality of DNA template influenced restriction enzyme digestion and the subsequent ligase amplification reaction, while the modified CTAB extraction method could be used in AFLP analysis of Chinese cabbage to obtain a clear AFLP fingerprint. The optimum conditions for restriction enzyme digestion of genomic DNA from Chinese cabbage were as follows: 150 g DNA template, 12.5 μl reaction volume, 1.25 U Eco R Ⅰ, 1.25 U Mse Ⅰ and 5×Reaction Buffer with 4 h at 37 ℃. The ligation reaction with 2.5 h at 20 ℃ was the optimum condition. Six pairs of primers including E-AAC/M-CAG, E-AAG/M-CAC, E-ACA/M-CTG, E-ACT/M-CAC, E-ACT/M-CTT and E-ACT/M-CTC all had its own stable and clear patterns. Conclusion] With abundant bands and high polymorphism, AFLP selective amplification is an efficient molecular marker for genomic polymorphism of Chinese cabbage.
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| 关 键 词： | 选择性扩增 提取 扩增产物 基因组 电泳图谱 双酶切 大白菜 引物组合 反应体系 |
Studies on Genomic DNA Extraction and Establishment of AFLP Reaction System in Chinese Cabbage |
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| MENG Shu-chun,ZHANG Hai-ying,ZHENG Xiao-ying,LIU Yu-mei,WANG Yong-jian. National Engineering Research Center for Vegetables,Beijing.Studies on Genomic DNA Extraction and Establishment of AFLP Reaction System in Chinese Cabbage[J].农业科学与技术(英文版),2009(3). |
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| Authors: | MENG Shu-chun ZHANG Hai-ying ZHENG Xiao-ying LIU Yu-mei WANG Yong-jian National Engineering Research Center for Vegetables Beijing |
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| Institution: | MENG Shu-chun1,ZHANG Hai-ying1,ZHENG Xiao-ying1,LIU Yu-mei2,WANG Yong-jian11. National Engineering Research Center for Vegetables,Beijing 100097,2. Institute of Vegetables , Flowers Chinese Academy of Agricultural Sciences,Beijing 100081 |
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| Abstract: | Objective] The obtained clear AFLP fingerprint of Chinese cabbage provided basis for studies on the molecular markers of Chinese cabbage cultivars and the phylogenetic relationship among Chinese cabbage cultivars. Method] With the test materials of leaves of Chinese cabbages, the high-quality total DNA from leaves of Chinese cabbages was extracted by the modified CTAB method. DNA restriction-ligase reaction, pre-amplification and selective amplification were optimized, and the AFLP silver-staining reactio... |
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| Keywords: | Chinese cabbage DNA extraction AFLP |
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