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《Northern Horticulture》 2008-04
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Optimization of RAPD Reaction systems for Curcuma wenyujin

WANG Xiao-hui1,TANG Xiao-chuang1,YANG En-xiu1,JIANG Cheng-xi1,LI Min2,LI Xiao-kun1,2(1.Ministry of Education,Engineer Research Center of Bioreactor and Pharmaceutical Development,Jilin Agricultural University,Changchun 130118,China;2.Pharmacy School,Wenzhou Medical College,Wenzhou,Zhejiang 325035,China)  
Some reaction condition was optimized by amplifying technology and method of PCR based on isolating genomic DNA from Curcuma wenyujin,to establish the best reaction condition of RAPD-PCR amplification system of Curcuma wenyujin.Results showed that the best amplification system for Curcuma wenyujin was as follows:total reaction volume 25 μL,including Mg2+(25 mM)2.0 μL,Taq polymerase(5 U/L)0.3 μL,random primer(20 M)1.0 μL,template DNA 1.0 μL,dNTP(2.5 mM)2.0 μL,10×PCR buffer 2.5 μL;the amplification procedure conditions were pre-denature at 94℃ for 5 min followed by denature at 94℃for 35 s,annealing at 36℃ for 1 min,extension at 72℃ for1.5 min,cycling 42 times,last extension at 72℃ for 10 min.This amplification system was very economical,convenient and the band was clear and stable.It was the most appropriate system,based for the genetic diversity of curcuma wenyujin.
【Fund】: 浙江省重大科技资助项目(No2005C13019);; 瑞安市重大科技资助项目(20052005);; 温州市科技计划资助项目(S2005B001)
【CateGory Index】: R285
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