Optimization of RAPD conditions for total microbes in fermented meat

The total microbial DNA was extracted by the improved benzyl chloride method from fermented meat and was analyzed by random amplified polymorphic DNA(RAPD) markers.The influence of the concentration of Mg2+,dNTPs,primer,Taq DNA polymerase and the templates on RAPD-PCR reaction system were investigated by single-factor gradient test method.The optimal conditions were as follows: MgCl2 concentration 2.5mmol/L,dNTPs concentration 0.15mmol/L,primer concentration 15pmol,DNA template 50ng and Taq DNA polymerase 1.0U in 25μL RAPD-PCR reaction system.