甜菜多重SSR-PCR体系的建立和优化

为了建立甜菜多重SSR-PCR体系，通过利用11对甜菜SSR核心引物，根据SSR扩增产物片段大小的不同，构建甜菜2~5重SSR-PCR反应体系。结果表明，在单一SSR-PCR的基础上，甜菜2~3重SSR-PCR的体系为：每增加一重SSR，仅仅增加相应引物的量以及减少去离子水的量。甜菜4~5重SSR-PCR，要在单一PCR的基础上增加0.5倍DNTPs的含量以及相应引物的量，同时根据个别引物扩增效率的不同，相应减少或者增加0.5倍个别引物的量，并成功的构建了16个4重PCR和9个5重PCR。多重PCR反应能够产生与单一PCR相同的多态性，但是却比单一PCR提高了2~5倍的效率，甜菜多重PCR体系的建立将大大加速甜菜品种纯度和真实性鉴定的速度，也将更快的促进甜菜分子生物学其他领域的发展。

Construction and Optimization of Multiplex PCR of Maize with SSR Markers

In order to build the multiple SSR-PCR system for sugar beet, the 11 pairs of sugar beet SSR core primer were introduced to build 2 fold to 5 fold SSR-PCR reaction system, based on the different fragment size of SSR amplification products. The results showed as follows: on the basis of single SSR-PCR, the 2 fold to 3 fold SSR-PCR system for sugar beet showed: each one fold increase in SSR, there was only quantity increase in corresponding primer and quantity reduced in deionized water. As to the 2 fold to 3 fold SSR-PCR system for sugar beet, on the basis of single PCR, 0.5 fold increased in DNTPs content quantity and corresponding primer quantity, meanwhile correspondingly reduce or increase 0.5 fold quantity for individual primer according to the different amplification efficiency of individual primer, then we could successfully build 16 4-fold PCR and 9 5-fold PCR. The polymorphism produced by multiple PCR reaction was identical with that from single PCR, but there showed a 2 fold to 5 fold higher efficiency than the single PCR. The establishment of multiple PCR system for sugar beet would greatly accelerate the speed for identifying the variety purity and authenticity of sugar beet, and further promote the development of other areas in sugar beet molecular biology.