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《Journal of Anhui Agricultural Sciences》 2008-03
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DNA Extraction from Camellia oleifera and the Establishment of the Optimum Reaction System of RAPD Analysis

QUE Sheng-quan et al(Department of Biology and Environmental Engineering,Nanchang University of Technology,Nanchang,Jiangxi 330013)  
[Objective] The research aimed to establish the optimum reaction system of RAPD analysis for Camellia oleifera.[Method] Genomic DNA was extracted from fresh tender leaves in 12 species of C.oleifera by using the improved SDS method and CATB respectively.The purity of DNA extracted by 2 kinds of methods was compared by determining the values of OD260 and OD280.The RAPD reaction procedure for C.oleifera was established,and the optimum reaction system of RAPD analysis for C.oleifera was obtained through optimizing the reaction conditions.[Result] The purity of genomic DNA extracted from C.oleifera by CTAB method was higher than that by SDS method.The optimum reaction system of RAPD analysis for C.oleifera was as follows: 1.0 U Taq DNA polymerase,1.5 mmol/L MgCl2,200 μmol/L dNTP,10 pmol/L random primers,20 ng template DNA,2 μl 10×PCR buffer,adding the redistilled water till 20 μl.The RAPD reaction procedure for C.oleifera was as follows: predenaturation at 95 ℃ for 3 min;denaturation at 94 ℃ for 1 min,annealing at 40 ℃ for 1 min,extension at 72 ℃ for 2 min,40 cycles and extension at 72℃ for 7 min.[Conclusion] The DNA used in this research had high purity,the reaction conditions were strictly controlled and the amplification stability in the test was better,which was the optimum reaction system of RAPD analysis for C.oleifera.
【CateGory Index】: S794.4
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