【摘要】 以罗汉果 ( Siraitia grosvenorii) DNA为材料 ,分析了模板 DNA,Mg2 + ,d NTPs,引物的浓度 ,Taq DNA聚合酶的用量以及循环次数对 ISSR-PCR扩增结果的影响 ,确立了稳定的、可重复的罗汉果 ISSR最佳反应体系和 PCR扩增参数 :在 2 5μL的 PCR反应体系中 ,含 2 0～ 5 0 ng模板 DNA,1 U Taq酶 ,1× PCR缓冲液 ,2 .0 mmol/L Mg Cl2 ,4种 d NTPs各 2 0 0 μmol/L,0 .5 μmol/L引物 ;PCR扩增程序为 94°C预变性 3 min,接着进行 40个循环 :94°C变性 1 min,5 2°C退火 5 0 s,72°C延伸 2 min,循环结束后 72°C延伸 7min.更多还原

【Abstract】 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,Mg²⁺,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 MgCl₂,200 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.更多还原