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

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