【摘要】 以四川泽泻为实验试材,采用改进的CTAB法提取了泽泻的高质量总DNA模板,克服了泽泻中所富含的酚类物质的影响.对泽泻ISSR反应体系中各个主要影响因子进行了优化和筛选,建立了标记位点清晰、稳定、重复性好、可用于泽泻ISSR-PCR分析的最适反应体系,它们是:25μL PCR反应体系中,10×Taq酶配套缓冲液,1U Taq DNA聚合酶,1.8~2.0 mmol/L MgC l2,100μmol/L dNTP,0.3μmol/L引物,DNA模板约10~20 ng,退火温度在52~60℃;为进一步研究泽泻居群的ISSR分子标记鉴别奠定了基础.更多还原

【Abstract】 Alisma orientalis（Sam.） Juzep of Szechwan is used as an experimental material.The improved method of CTAB is adopted to distill total DNA template of high quality and the infection that Alisma orientalis contains abundant kinds of hydroxybenzene has been conquered.ISSR-PCR reaction effects are tested and optimized.The optimal ISSR-PCR reaction system in Alisma orientalis is established: 25μL PCR reaction volume,10×Taq buffer,1.5U Taq DNA polymerase, 1.8-2.0mmol/L MgCl₂,100μmol/L dNTP,0.3 μmol/L primer and 10-20ng template DNA.The appropriate annealing temperature is among 52-60℃.This article provides clear,reliable,abundant polymorphisms molecular markers suitable for authenticating the populations of Alisma orientalis.更多还原