Optimizing System of SSR-PCR in Peach

摘要

In order to get more in-depth studies of germplasm identification, germplasm resources research, genetic linkage map drawing marking the orientation of the characteristic gene and molecular-assisted breeding and so on , a stable, efficient and reproducible PCR system need to be founded. So a L16(45) (DNA template, Taq DNA polymerase, 10 x Buffer (Mg2T) , primer, and dNTPs) orthogonal design was used to optimize SSR-PCR amplification system using peach genomic DNA as template, then through the reaction efficiency which was affected by these factors determined the best one. Last, two methods were used to verify its stability. Based on the results, a stable, efficient and reproducible 20uL PCR system were obtained, it containing Taq DNA polymerase 1.2U, 10 x Buffer (Mg2) 4uL, lOmmolL-1 dNTPs 0.7uL, 5mmolLl SSR primer0.8μL, DNA template 40 ng .