论文部分内容阅读
将毛细管电泳四色荧光检测技术应用于茶树SSR标记分析.该方法采用三引物PCR扩增SSR位点,三引物即在5′端加有M13尾巴序列(5′-CACGACGTTGTAAAACGAC-3′)的特异正向引物、特异反向引物及带有荧光标记的通用型M13引物;为了运用四色荧光检测系统使通过一次毛细管电泳能同时检测3个以上的SSR位点,采用蓝、绿、黑3种不同颜色的荧光染料分别对3个M13引物进行标记.应用该方法对42个茶树品种(系)的16个SSR位点进行遗传分析的结果表明:此法具有简便、可靠、低成本及高通量的优点;且随着所分析SSR位点数的增加,降低成本的效果更加显著.采用建立的方法,还筛选获得了11个多态性丰富的可应用于茶树遗传研究的SSR标记.
The four-color fluorescence detection technique of capillary electrophoresis was applied to the SSR marker analysis of tea plant.This method uses three primers to amplify the SSR locus, and the three primers are specific to the 5 ’end of M13 tail sequence (5’-CACGACGTTGTAAAACGAC-3’) Forward primer, specific reverse primer and universal M13 primer with fluorescent marker. In order to use the four-color fluorescence detection system to simultaneously detect more than three SSR sites by one capillary electrophoresis, three types of blue, green and black Three M13 primers were labeled with fluorescent dyes of different colors respectively.The genetic analysis of 16 SSR loci in 42 tea cultivars (lines) using this method showed that this method was simple, reliable, low cost and high-pass And the cost reduction was more significant with the increase of the number of SSR loci analyzed.We also screened out 11 SSR markers rich in polymorphism that can be applied to the genetic research of tea plant by using the established method.