TP-M13自动荧光法的改进及在美国山核桃品种鉴定上的应用

Modified TP-M13 Automated Fluorescent-labelled SSR Method and Its Application in Identification of Pecan Cultivars

摘要: 用TP-M13自动荧光法检测了常用10个美国山核桃品种中的8个SSR位点，发现原有直接使用三引物方法扩增的有效性和可靠性较低，而经过改进的间接三引物法则有较大的实用价值，在此基础上进一步研究后认为混合三引物法在适当控制混合引物的对数和提高特异性后，在提高间接三引物法的扩增效率方面是可行的。

Abstract: Although TP-M13 automated fluorescent-labelled system has reduced the cost of fluorescent detection of SSR genotyping by introducing a universal fluorescent labelled primer, the complexity of PCR reaction brought by three primers limits its application on more species. TP-M13 method using three primers directly was implemented to detect eight SSR loci of regular 10 pecan (Carya illinoensis ) cultivars, which revealed the low availability and liability. A modified TP-M13 detection method called indirect three primer method was proved to be effective and valuable in solving this problem. Furthermore, the mixing of SSR primers based on this indirect three primer method could further improve the PCR efficiency