中国玉米新品种DNA指纹库建立系列研究Ⅲ.多重PCR技术在玉米SSR引物扩增中的应用

根据21对玉米SSR引物扩增片段大小范围组成不同的引物组合进行多重PCR反应的研究。在与单一PCR相同的反应条件下，47个两重PCR组合出现5种不同的扩增情况，其中30个组合扩增正常；10个三重PCR组合中有9个组合扩增正常；两个四重PCR组合中有1个扩增正常，这些筛选出的正常扩增的组合可应用于今后玉米DNA指纹库构建研究中。研究表明，应用与单一PCR完全相同的反应条件，大部分能够获得正常扩增的多重PCR组合，在此基础上提出了简化的多重PCR优化程序。

Series of Research on Establishing DNA Fingerprinting Database of Chinese New Maize Cultivars Ⅲ. Multiplex PCR Applied in Maize SSR Amplification

Twenty-one pairs of maize SSR primers were used for building various multiplex-PCR-sets according to their amplified bands'size range.Under the same conditions as single PCR reaction,47 two-PCR-sets appeared five different amplification behaviors,of which 30 were amplified normally.Nine of 10 three-PCR-sets and one of 2 four-PCR-sets were amplified normally.These screened normally-amplified multiplex-PCR-sets could be applied in maize DNA fingerprinting database.The results showed that most multiplex-PCR-sets could be amplified normally under just the same conditions as single PCR.Based on our experience,we proposed a simolified protocol for developing a multiplex PCR assay.