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《Northern Horticulture》 2008-05
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Establishment of Eustoma grandiflorum Optimal RAPD Reaction System Using Orthogonal Design

WANG Yi,CHEN Chong-shun,CAO Wei-jie,WANG Zhuan-mei(Jiangsu Key Laboratory for Biodiversity & Biotechnology,College of Life Sciences,Nanjing Normal University,Nanjing 210046,China)  
The essential factors of the RAPD reaction system for Eustoma grandiflorum,such as:Mg2+,dNTPs,Taq DNA polymerase,primer and DNA template were optimized by using the method of orthogonal design.The optimal RAPD reaction system was set as follows: 10×PCR buffer 2.5 μL,Mg2+ 2.0 mmol/L,dNTPs 0.4 mmol/L,random primers 0.2 μmol/L,DNA template 50 ng/μL and Taq DNA polymerase 1.0 U,in a total 25 μL reaction volume.Then the effects of different annealing temperatures to the amplified products of RAPD were also investigated.The annealing temperature was selected as 41℃.Finally the optimized system was successfully applied to different random primers,indicating that the optimal system was stable and adapted.
【CateGory Index】: S682.19
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