Establishment of multiplex-PCR for waxy genes in wheat.

The research studied the influences of PCR components and cyclic parameters on the multiplex PCR and established a multiplex PCR system to simultaneously identify Wx-A1、Wx-B1 and Wx-D1 genes of wheat in one reaction.The result showed that the primers concentration ratio and Tm value were the critical factors to influence effectiveness of multiplex PCR,when the primers concentration(SSR/MAG267)was 1(0.1 μmol/L∶0.1 μmol/L)~1.22(0.11 μmol/L∶0.09 μmol/L)and Tm=58℃,the outcome of multiplex PCR of Wx-A1,Wx-B1 and Wx-D1 was the best.The multiplex PCR system identifying three Wx genes quickly and efficiently simultaneously in one reaction can be used to carry out multiplex molecular marker-assisted selection in breeding waxy wheat cultivars and quality wheat breeding.