不同PCR程序对甜菜DAMD引物扩增效果的影响

Effect of Different PCR Programs on DAMD Primer Amplification in Sugar Beet

为了开发甜菜品种高效鉴定的DAMD-PCR分子标记技术,优化相应的检测技术流程。利用6种DAMD引物,以8个不同甜菜品种的DNA为模板,分别采用常规PCR扩增程序和touchdown PCR(TD-PCR)扩增程序对模板进行扩增。扩增产物利用8%的非变性聚丙烯酰胺凝胶电泳进行分离,经银染后观察不同PCR程序对甜菜DAMD引物扩增效果的影响。结果表明,TD-PCR扩增程序扩增效果明显好于常规PCR扩增程序,条带明亮、清晰,非特异性条带数量明显减少。推荐在甜菜DAMD引物的扩增中使用TD-PCR扩增程序。

In order to develop the DAMD-PCR molecular marker technology for high efficiency identification of sugar beet varieties, the corresponding detection process was optimized. A total of 6 DAMD primers were selected for amplification of DNA in 8 different sugar beet varieties based on both standard and touchdown PCR programs. The PCR products were separated by native PAGE. After silver stain, the effects of DAMD-PCR amplification by different PCR programs were analyzed. The results suggested that touchdown PCR had better amplification efficiency than that of standard PCR program, and the bands were bright, clear and the number of non-specific bands was significantly reduced. It is recommended for DAMD-PCR in term of sugar beet varieties identification.