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《Chinese Journal of Tropical Crops》 2009-11
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Optimization of ISSR-PCR in Cynodon dactylon

Huang Chunqiong1,2,3,Liu Guodao1,2 1 Tropical Crop Genetic Resources Institute,CATAS,Danzhou,Hainan 571737;2 Ministry of Agriculture Key Laboratory for Utilization of Tropical Crop Germplasm Resources,Danzhou,Hainan 571737,China 3 College of Agronomy,Hainan University,Danzhou,Hainan 571737  
The concentrations of Mg2+,dNTPs,Taq DNA polymerase,primers and DNA template which affect the ISSR-PCR reactions were optimized by using an orthogonal design with 5 factors at 4 levels.Results showed that the optimum system for 25 μL ISSR-PCR reaction was Mg2+ 2.5 mmol/L,dNTPs 0.25 mmol/L,primer 0.2 μmol/L,Taq DNA polymerase 1.0 U,template DNA 40 ng and that the optimal annealing temperature of primer UBC881 was 50.5 ℃.Under this optimum system amplifications carried out on 24 genotypes of Cynodon dactylon(Linnaeus) Persoon gave better results.The system was steady and reliable and could be applied to molecular genetic research of C.dactylon.
【Fund】: 热带牧草种质资源保护项目;; 国家科技支撑计划(No.2008BADB3B08)项目;; 973项目(No.2007CB108903);; 中国热带农业科学院热带作物品种资源研究所研究生基金项目(No.YJS-2008-B002)资助
【CateGory Index】: S688.4
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