摘要
利用正交设计L16(45)方法对迎红杜鹃(Rhododendron mucronulatum Turcz.)ISSR-PCR(Inter-simple sequence repeat-Polymerase chain reaction products)反应体系的5因素(TaqDNA聚合酶、Mg2+、DNA模板、dNTP、引物)在4个水平上进行了优化试验。结果表明,ISSR-PCR反应体系各因素在设定的不同水平对PCR反应结果都有影响,其中引物浓度的影响最大;筛选出了各反应因素的最佳水平,建立的迎红杜鹃ISSR-PCR反应最佳体系(25μL)为2.0μL10×Buffer、0.3μL Taq DNA聚合酶(5U/μL)、2μLDNA模板(20ng/μL),2.5μL dNTPs(2.5mmol/L)、2μL引物(10μmol/L)。
The orthogonal design was used to optimize Rhododendron mucronulatum Turcz.＇s ISSR-PCR（Inter-simple sequence repeat-Polymerase chain reaction products） amplification system＇s five factors（Taq DNA polymerase,Mg2＋,DNA template,dNTP,and primer） in four levels respectively.The results showed that,the affections of each factor in different levels were found,and concentrations of primer was the most effective factor to the result of PCR,a most suitable ISSR-PCR system for R.mucronulatum was established,it＇s 25 μL reaction system which contained 2.0 μL 10×Buffer,0.3 μL Taq DNA polymerase（5 U/μL）,2 μL DNA template（20ng/μL）,2.5 μL dNTPs（2.5 mmol/L）,and 2 μL primer（10 μmol/L）.
出处
《湖北农业科学》
北大核心
2010年第11期2657-2659,共3页
Hubei Agricultural Sciences
关键词
迎红杜鹃
正交设计
简单重复序列区间-多聚酶链式反应体系
Rhododendron mucronulatum Turcz.
orthogonal design
Inter-simple sequence repeat-Polymerase chain reaction products system
