| 马铃薯SSR-PCR体系的优化与建立 |
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| 引用本文： | 李建武,李高峰,胡新元,文国宏,魏子杰. 马铃薯SSR-PCR体系的优化与建立[J]. 中国马铃薯, 2013, 0(6): 331-335 |
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| 作者姓名： | 李建武 李高峰 胡新元 文国宏 魏子杰 |
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| 作者单位： | [1]甘肃省农业科学院马铃薯研究所,甘肃兰州730070 [2]农业部西北旱作马铃薯科学观测实验站,甘肃渭源748201 [3]甘肃省农业科学院生物技术研究所,甘肃兰州730070 [4]甘肃农业大学生命科学技术学院,甘肃兰州730070 |
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| 基金项目： | 国家自然基金“四倍体马铃薯分子遗传图谱的构建与淀粉含量等重要农艺性状的QTL定位研究”（31160299）. |
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| 摘 要： | 以马铃薯叶片DNA为模板,采用单因素试验的方法,对影响马铃薯SSR-PCR反应体系的主要成分模板DNA、dNTP浓度、引物浓度、Mg2+浓度、Taq酶浓度及退火温度进行了优化,并建立最优化的马铃薯SSR扩增反应体系。结果表明:在20μL反应体系中,模板DNA用量为60 ng,dNTP为0.3125 mmol/L,引物浓度为2.4pmol,Mg2+为1.5 mmol/L,Taq酶为0.5 U,筛选各引物最适宜的退火温度,经2.0%琼脂糖凝胶电泳检测,扩增条带清晰,优化后的SSR反应体系稳定性好,可用于马铃薯遗传多样性分析。
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| 关 键 词： | 马铃薯 SSR—PCR 优化 |
Establishment and Optimization of SSR-PCR Reaction System of Potato |
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| LI Jianwu,LI Gaofeng,HU Xinyuan,WEN Guohong,WEI Zijie. Establishment and Optimization of SSR-PCR Reaction System of Potato[J]. Chinese Potato, 2013, 0(6): 331-335 |
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| Authors: | LI Jianwu LI Gaofeng HU Xinyuan WEN Guohong WEI Zijie |
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| Affiliation: | 1. Institute of Potato, Gansu Academy of Agricultural Sciences, Lanzhou, Gansu 730070, China; 2. Scientific Observing and Experimental Station of Potato Dry Farming in Northwest China, Ministry of Agriculture, Weiyuan, Guansu 748201, China 3. Institute of Biotechnology, Gansu Academy of Agricultural Sciences, Lanzhou, Gansu 730070, China 4. College of Life Science and Technology, Gansu Agricultural University, Lanzhou, Gansu 730070, China ) |
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| Abstract: | The main component which affects SSR-PCR reaction system on potato, including template DNA, dNTPs concentration, primer concentration, buffer concentration, Taq enzyme concentration and annealing temperature, were optimized in a single factor experiment, and the optimized SSR amplification reaction system was established. The results indicated that 20 μL volume SSR-PCR system containing 60 ng template DNA, 0.3125 mmol/L dNTPs, 2.4 pmol primer, 1.5 mmol/L Mg2＋, and 0.5 U Taq polymerase, combined with optimal annealing temperature for each primer, performed wel , and the optimized system of SSR-PCR, which was steady and reproducible, could be applied to the study of genetic diversity of potato after PCR results were run by 2.0%agarose gel. |
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| Keywords: | potato SSR-PCR optimization |
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