High-throughput S-SAP by fluorescent multiplex PCR and capillary electrophoresis in plants |
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Authors: | Tang Tian Huang Jianzi Zhong Yang Shi Suhua |
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Institution: | State Key Laboratory of Biocontrol, School of Life Sciences, Zhongshan University, Guangzhou 510275, Guangdong, PR China. |
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Abstract: | The inherent replicative mode of transposition endows retrotransposons with considerable advantages as genetic tools in plant genome analysis. Here we present a high-throughput sequence-specific amplification polymorphism (S-SAP) method based on copia-like retrotransposons to fulfill the increasing desire of screening large numbers of samples in plants. Classic approach for digestion, ligation and pre-amplification was combined with optimized fluorescent multiplex PCR for simultaneously selective amplifying S-SAP fragments, and multiple S-SAPs were subsequently detected by capillary electrophoresis using ABI PRISM 3700 capillary instruments. Comparisons of results from multiplex PCR with simplex PCR, and from capillary electrophoresis with slab-gel electrophoresis demonstrated that this method is an efficient, economical, and accurate means for high-throughput and large-scale genotyping retrotransposon variation in plants. |
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