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当前位置: 首页 > 文章 > 整粒和半粒青稞种子DNA的提取及RAPD检测 西南农业学报 2017 (6) 1257-1261
Position: Home > Articles > DNA Extraction and RAPD Detection of Half-grain and Whole-grain Hordeum vulgare var. nudum Southwest China Journal of Agricultural Sciences 2017 (6) 1257-1261

整粒和半粒青稞种子DNA的提取及RAPD检测

作  者：
郝豆豆;张勇群;高玉花;雷鸣;武俊喜;拉多
单  位：
西藏自治区人民政府驻成都办事处医院;中国科学院地理科学与资源研究所;西藏大学
关键词：
青稞种子;基因组DNA;RAPD-PCR扩增
摘  要：
【目的】从青稞种子中提取高质量的基因组DNA。【方法】以半粒无胚青稞种子和整粒青稞种子为材料,用6种不同的方法提取基因组DNA,用琼脂糖凝胶电泳检测DNA的完整性,紫外分光光度计法检测DNA的浓度和纯度,并对提取的DNA进行了RAPD-PCR扩增检测,同时将剩下的半粒有胚青稞种子进行萌发实验。【结果】从半粒和整粒青稞种子中都可以提取到高质量的DNA,用提取的DNA作为RAPD-PCR扩增检测模板,得到的扩增产物条带清晰,多态性条带丰富,其质量满足RAPD分子标记的要求。【结论】高盐低p H法提取的DNA质量最好,是这6种方法中的最佳方法;半粒青稞有胚种子也能够正常生根发芽,可作为遗传实验的材料。
译  名：
DNA Extraction and RAPD Detection of Half-grain and Whole-grain Hordeum vulgare var. nudum
作  者：
HAO Dou-dou;ZHANG Yong-qun;GAO Yu-hua;LEI Ming;WU Jun-xi;LHA-duo;Hospital of Chengdu Office of People's Government of Tibetan Autonomous Region;Tibet University;Institute of Geographical Sciences;
关键词：
Hordeum vulgare var;;nudum seed;;Genomic DNA;;RAPD-PCR amplification
摘  要：
【Objective】The high-quality DNA could be extracted from Hordeum vulgare var. nudum. 【Method】The genomic DNA was extracted by six different methods from half-grain embryoless seeds and whole-grain seeds of Hordeum vulgare var. nudum,UV Spectrophotometry and agarose gel electrophoresis were used to detect the concentration,purity and completeness of DNA. RAPD-PCR amplification was carried out on the extracted DNA. At the same time,the left half-grain embryo seeds were cultured for germination experiments. 【Result】High-quality DNA was able to be extracted from both half-grain and whole-grain Hordeum vulgare var. nudum seeds. The extracted DNAs were employed as the template for RAPD-PCR amplification detection,and the products of amplification have clear and rich polymorphism bands,the quality of extracted DNA could meet the requirement of RAPD molecular marker experiment. 【Conclusion】High salt and low p H which could extract high quality DNA is the best method of six methods. In addition,the sprouting experiments indicated that the left halfembryo seed was able to sprout normally and can be used as material of genetic experiment.

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