摘要
利用正交试验设计法L9(34),从引物浓度、TaqDNA聚合酶浓度、Mg2+浓度和dNTP浓度4种因素3个水平,对葛资源ISSR-PCR反应体系进行优化分析,并在此基础上对PCR反应的退火温度进行梯度筛选。结果表明,25μL最佳的葛资源ISSR-PCR反应体系是:1×PCR buffer、0.20 mmol/L dNTP、0.4μmol/L引物、2.5 mmol/L Mg2+和0.5 U TaqDNA聚合酶,引物UBC809的最佳退火温度为57.9℃。
In this study, orthogonal experimental design method L9 （34）was used, from the concentration of primers, Taq DNA polymerase concentrations, Mg2＋ concentration and dNTP concentrations of four kinds of factors in three levels, ISSR-PCR reaction system of Pueraria was analysed. On this basis, the annealing temperature were proposed by gradient PCR. The results showed that, 25 μL ueraria resources ISSR-PCR reaction system was： lxPCR buffer, 0.20 mmol/L dNTP, 0.4 μ mol/L primer, 2.5 mmol/L Mg2＋and 0.5 U Taq DNA polymerase, the optimal annealing temperature of primer UBC809 was 57.9℃.
出处
《广东农业科学》
CAS
CSCD
北大核心
2010年第1期120-123,共4页
Guangdong Agricultural Sciences
基金
江西省教育厅科技项目(GJJ09413)
关键词
野葛
粉葛
ISSR—PCR
正交优化
Pueraria lobata （Willd） Ohwi
Pueroria thomsonii Benth
ISSR-PCR
orthogonal optimization
