青枯菌SRAP-PCR体系的建立与优化

Establishment and Optimization of SRAP Analysis System of Ralstonia solanacearum Genome

本研究以马铃薯青枯菌P046为试验材料,应用单因素试验法优化SRAP-PCR反应体系,得出重复性好、带型清晰、稳定性强的最佳反应体系:总体积为10μL,50 ng/μL模板DNA 0.6μL、10μmol/L引物0.8μL、Tap DNA聚合酶1.0 U、10 mol/L、dNTPs 1.0μL.该反应体系可用于青枯菌遗传多样性、基因定位与克隆等的研究.

Some factors affecting PCR amplification of SRAP（Sequence-related amplified polymorphism） were studied using Ralstonia solanacearum P046. A reliable, effective,reproductive and legible repeats optimized technical system was developed. In the 10 μL PCR reaction nfixture,30 ng of genomic DNA, 1.0 U of Taq polymerase,0.8 μL 10 umol/L primers,and 1.0 μL dNTPs were used. This reaction system can be used to study genetic diversify,gene tagging cloning and genetics mapping for SRAP of differ- cnt Ralstonia solanacearum.