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《Northern Horticulture》 2009-02
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Optimization for RAPD-PCR Reaction System and Amplified Program of Dry Peppers

WANG Wei1,ZHENG Gen-chang1,YAN Guang-zhao1,LIU Peng1,CHEN Bing-yan1,BAO Feng-li2(1.Inner Mongolia University for Nationalities,Tongliao,Inner Mongolia 028043,China;2.Denong Need Co.LTD,Beijing 100101,China)  
The factors influencing RAPD analysis,including Taq polymerase,Mg2+,dNTPs,primers,template DNA concentration,denaturing time and thermal cycles in Dry peppers were studied.An optimal PCR system for RAPD in fresh leaves of Dry peppers had been found in 25 μL reaction solution,contained Taq polymerase1.5 U,Mg2+ 2.5 mmol/L,dNTPs 0.6 mmol/L,primers 0.8 μmol/L,template DNA 60 ng.The amplification program was devised:initial denaturing at 94℃ for 4 min;denaturing at 94℃ for 1 min,annealing at 37℃ for 1 mins,extension at 72℃ for 1.5 min,40 cycles;final extension at 72℃ for 5 mins.Steadily and realibly profile was obtained in the dry Pepper using the optimal system and amplification program,basic job for studing dry pepper genetic diversity.
【CateGory Index】: S641.303.6
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