稻瘟病菌SSR检测是分子标记辅助育种的一项重要技术。为优化SSR反应体系，以稻瘟病菌菌株504为供试材料，采用单因素筛选法及L9(34)正交试验设计，研究了稻瘟病菌SSR分析中PCR反应体系的主要成分对扩增结果的影响。结果表明：在总体积为20μL的PCR反应中，Taq DNA聚合酶的最适用量为1.0U；Mg2+、dNTP和引物的最适终浓度分别为1.0mmol/L、100μmol/L和0.4μmol/L。利用该体系进行扩增，所得谱带清晰、稳定、非特异性带少。
Abstract
SSR detection is an important technique of molecular marker for assisted breeding in Pyricularia grisea. In order to optimize SSR-PCR reaction system, the main factors affecting amplication results were studied with single factor selection and L9(34) orthogonal design. The results showed that the optimal reaction system was as follows： the 20μL reaction system contained TaqDNA polymerase 1.0U, Mg2+ 1.0 mmol/L, dNTP 100 μmol/L and Primer 0.4μmol/L. Using this system, amplication bands were clear, standard and non-especial bands were shorter.
关键词
稻瘟病菌；SSR；体系优化
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Key words
Pyricularia grisea;SSR;System optimization
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参考文献
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