西瓜枯萎病菌最优RAPD体系的建立

Establishment of RAPD technique system on Fusarium oxporum f.sp.niveum

采用完全随机试验,对适合西瓜枯萎病菌的最优RAPD体系(包括随机引物浓度、模板DNA浓度、TaqDNA聚合酶浓度以及扩增反应中的退火温度)进行了探索,寻求各因素间的最优组合,最终得到了条带清晰的最优RAPD体系。反应体积为25μL,其中随机引物浓度为10μmol L,TaqDNA聚合酶为1 25U,模板DNA浓度为1 6mg L,dNTPs浓度为2mmol μL,扩增反应的退火温度为37℃。

Random thoroughly experiment was made to investigate the optimization of RAPD technique system(including the concentration of random premier, template DNA and Taq DNA polymerase, and annealing tempreture in the amplification reaction), seek the best combination among factors. A satisfactory RAPD technique system for the pathogen DNA with clear bands was established.The concentration of random premier was 10 μmol/L, template 1.6 mg/L, Taq DNA polymerase 1.25 U, dNTPs 2 mmol/μL, and the annealing tempreature 37℃.