人参基因组DNA的提取及RAPD—PCR反应条件的优化

探讨了人参基因组DNA提取方法及RAPD—PCR反应条件的优化．结果表明，用改良的CTAB方法提取的DNA均达到了RAPD—PCR分析的要求；优化的RAPD—PCR反应条件为：在25μLRAPD—PCR反应体系中，模板DNA20ng，dNTP200μmol／L，MgCl2 1．5mmol／L，10×Buffer2．5μL，引物浓度0．4pmol／L，TaqDNA聚合酶1．0U，其余以ddH20定容至25μL．PCR反应程序为：94℃预变性5min；94℃45S，40℃退火1min，72℃1min，共40个循环；最后在72℃延伸5min．

Genomic DNA extraction and optimization of RAPD-PCR conditions of Ginseng

Investigated the method of genomic DNA extraction and optimal RAPD--PCR condition. The results showed that the DNA were suitable for RAPD--PCR analysis extrac- ted by CTAB method with some variant;the optimal RAPD--PCR contitions is as follow： in total 25 μL volumes of PCR reaction,containing 20 ng of DNA,200 μmol/L of dNTPs, 1.5 mmol/L of Mge＋ ,2.5 μL of 10× buffer,0.4 μmol/L of primer, 1.0 U of Taq DNA polymerase. The PCR program is：after predenaturing at 94 ℃ for 5 min,followed by 40 cy- cles of denatureing at 94 ℃ for 45 s,annealing at 40 ℃ for 1 min,extension at 72 ℃ for lmin,and final extension at 72 ℃ for 5 min.