Factors including Mg^2＋, dNTP and primer that affected RAPD were studied using orthogonal experimental design, and at the same time, anneal temperature, extending time and cycles were also studied. Finally, PCR react...Factors including Mg^2＋, dNTP and primer that affected RAPD were studied using orthogonal experimental design, and at the same time, anneal temperature, extending time and cycles were also studied. Finally, PCR reaction system that is feasible for black soil microbial PCR amplification was determined, containing 7 ng DNA template, 20 pM random primers, 1.5 U Taq DNA polymerase, 3.0 mM MgCl2 and 0.2 mmol ·L^-1 dNTP, with procedure： 40 cycles for 3 min at 94 ℃ （temperature）, 40 s at 37 ℃ （annealing）, 1.5 min at 72 ℃ （extension）, a final elongation step at 72 ℃ for 7 min.展开更多
基金This Work was Supported by the National Natural Science Foundation of China (30230250, 30370819) Heilongjiang ProvinceScience Foundation (C0215)
文摘Factors including Mg^2＋, dNTP and primer that affected RAPD were studied using orthogonal experimental design, and at the same time, anneal temperature, extending time and cycles were also studied. Finally, PCR reaction system that is feasible for black soil microbial PCR amplification was determined, containing 7 ng DNA template, 20 pM random primers, 1.5 U Taq DNA polymerase, 3.0 mM MgCl2 and 0.2 mmol ·L^-1 dNTP, with procedure： 40 cycles for 3 min at 94 ℃ （temperature）, 40 s at 37 ℃ （annealing）, 1.5 min at 72 ℃ （extension）, a final elongation step at 72 ℃ for 7 min.
