Optimization of ISSR-PCR Reaction System and Primer Selection of Sophora tonkinensis Gagnep
Taking Sophora tonkinensis as materaial,orthogonal experiments were used to investigate the effects of DNA,primers,Mg2+,dNTPs,and Taq enzymes on the ISSR-PCR reaction system,and the conditions of ISSR-PCR reaction system of Sophora tonkinensis were es-tablished and optimized. The results showed that the optimal conditions for the ISSR-PCR reaction system (20μL) of Sophora tonkinensis in-cluding 3 mmol/L Mg2+,0.2 mmol/L dNTPs,1 U Taq enzyme,0.5μmol/L primer,100 ng template DNA. Ten primers with good polymor-phism were selected from 100 UBC primers by this system. The optimal system of ISSR-PCR reaction can be effectively established by orthogo-nal experiment design,which provides a scientific basis for the research of genetic relationship,genetic diversity evaluation of germplasm re-sources,variety identification and systematic classification of Sophora tonkinensis.
Sophora tonkinensis GagnepISSR-PCR system optimizationOrthogonal test designPrimer screening
万方数据
维普
