摘要
利用正交试验对影响RAPD-PCR反应体系的各种因素进行研究,从而建立适合杜鹃花RAPD反应的最佳反应体系,最终选择杜鹃基因组RAPD-PCR的最佳扩增反应体系,即模板DNA80ng、随机引物0.5μmol/L、dNTPs1.1mmol/L、MgCl22.5mmol/L,1.2μmol/LTag—DNA聚合酶,buffer2μL,总反应体系20μL。试验设计的6个因子、5个水平中,Mg2+、dNTPs和引物的浓度是主要因子,其中Mg2+的浓度对试验的影响最大,dNTPs和引物浓度在较低的水平就可满足需要,高浓度并没有显著提高试验效果。
Essential factors affecting the result of Rhododendron RAPD-PCR were studied with orthogonal design. And the optimal RAPD-PCR were built as follow：80ng DNA template, 0. 4μmol/L Random primer,1.1mmol/IANTPs,2. 5 mmol/LMgCl2and 0. 9μmol/L in 20gL reaction solution. By analyzing the 6 factors and 5 levels, the results showed that the concentration of Mg^2＋ , dNTPs and Random primer were the main factors,the concentration of Mg^2＋ had the greatest effect on the experiment. Low concentration of dNTPs and random primer ean reach the requirement.
出处
《北方园艺》
CAS
北大核心
2007年第5期124-126,共3页
Northern Horticulture
