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《Journal of Guangxi Normal University(Natural Science)》 2004-03
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ESTABLISHMENT OF ISSR-PCR SYSTEM IN LUOHANGUO (SIRAITIA GROSVENORII)

ZHOU Jun-ya,BIN Xiao-yun,PENG Yun-tao,TANG Shao-qing (College of Life Sciences,Guangxi Normal University,Guilin 541004,China)  
In this paper,the influencing factors of inter-simple sequence repeat (ISSR) were analyzed by using genomic DNA of Luohanguo(Siraitia grosvenorii).By adjusting the concentration of template DNA,Mg~(2+),dNTPs and primer,the contents of Taq DNA polymerase and the number of cycles,the stable and reproducible optimum reaction system of ISSR-PCR amplification and PCR amplification parameters were established for Luohanguo.The optimal system was in 25 μL reaction volume containing 20~50 ng template DNA,1 U Taq polymerase,1×PCR Buffer,2.0 mmol/L MgCl_2,200 μmol/L dNTPs and 0.5 μmmol/L primer.The amplification program was after 1 cycle initial denaturation at 94 °C for 3 min,each 40 cycles consisted of 94 °C 1 min,52 °C 50 s,72 °C 2 min,then final extension was at 72 °C for 7 min.
【Fund】: 国家自然科学基金资助项目 ( 3 0 2 60 0 1 3 )
【CateGory Index】: S668.9
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