| 藏獒基因组DNA提取及RAPD扩增体系的优化 |
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| 引用本文： | 兰小平,李三相,雒林通,李一婧,王廷璞,李燕,徐丽,鄢殉,崔泰保.藏獒基因组DNA提取及RAPD扩增体系的优化[J].湖北农业科学,2010,49(3). |
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| 作者姓名： | 兰小平 李三相 雒林通 李一婧 王廷璞 李燕 徐丽 鄢殉 崔泰保 |
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| 作者单位： | 1. 天水师范学院生命科学与化学学院,甘肃天水,741000
2. 甘肃农业大学,兰州,730070 |
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| 基金项目： | 甘肃省科学研究基金资助项目 |
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| 摘 要： | 采用改进的盐析法提取了藏獒血液基因组DNA,并对RAPD扩增体系进行了优化。结果表明,改进的盐析法所提取的藏獒基因组DNA完整、无降解,完全可以满足RAPD分析的需要;20μL的最佳扩增体系组成为2.50 mmol/L Mg2+、0.60 mmol/L dNTP、50 ng模板DNA、1.25U Taq酶和0.3μmol/L随机引物,最佳退火温度为36.5℃。
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| 关 键 词： | 藏獒 DNA提取 随机扩增多态性DNA 体系优化 |
Genomic DNA Extraction and Optimization of RAPD Reaction System of Tibetan Mastiff |
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| Abstract: | The method of improved salting out was used for whole blood genomic DNA extraction of Tibetan mastiff, and theRAPD reaction system of Tibetan mastiff was optimized. The results showed that the integrity and non-degradable genomicDNA of Tibetan mastiff could be gained by the method of improved salting out, and a total volume of 20 μL optimal RAPDreaction system of Tibetan mastiff consisted of 2.50 mmol/L Mg~(2+), 0.60 mmol/L dNTP, 50 ng genomic DNA, 1.25 U TaqDNA polymerase, and 0.3 μol/L arbitrary primers. The best renaturation temperature of RAPD analysis of Tibetan mastiffwas 36.5℃. |
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| Keywords: | Tibetan mastiff DNA extraction random amplified polymorphic DNA (RAPD) system optimization |
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