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《Journal of Anhui Agricultural Sciences》 2007-27
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Optimization of SRAP-PCR Program and System in Citrullus lanatus

LI Xiao-hui et al(Institute of Horticulture,Henan Academy of Agriculture sciences,Zhengzhou,Henan 450002)  
The aim of the research was to establish SRAP-PCR amplification system which was suitable for Citrullus lanatus DNA so as to lay foundation for the construction of gene map and molecular marker in C.lanatus.Watermelon variety-Zhonghuaquanwang being taken as tested materials,SRAP-PCR reaction program in watermelon was investigated and the gradient experiment was conducted on each impact factors in SRAP-PCR reaction system to screen and establish the optimum SRAP-PCR reaction program and system that could amplify high polymorphism,good repeatability and clear band pattern.The optimum SRAP-PCR reaction procedure was as follows:pre-denaturation at 94 ℃ for 5 min;denaturation at 94 ℃ for 1 min,anneal at 35 ℃ for 1 min,extension at 72 ℃ for 1 min and in total 5 cycles;denaturation at 94 ℃ for 1 min,anneal at 50 ℃ for 1 min,extension at 72 ℃ for 1 min and in total 35 cycles;extension at 72 ℃ for 5 min;preservation at 4 ℃.The optimum SRAP-PCR system(Total of 15 μl) was as follows: DNA 100 ng,dNTPs 0.3 mmol/L,Mg2+2.0 mmol/L primer 7.5 pmol/L,Taq polymerase 0.75 U.The program and system could meet the demands for genome SRAP amplification in C.lanatus.It was feasible to apply SRAP marker in genetic research in C.lanatus.
【Fund】: 河南省自然科学基金项目(0611031800)
【CateGory Index】: S651
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