本文探讨了红松(Pinus koraiensis Sieb. et Zucc.) ISSR实验中的各种影响因素,分别对红松的鲜叶、干叶、胚和胚乳4种材料进行了DNA的提取和PCR扩增,证明4种取样方式都是可行的。另外分析了模板的纯度和浓度、dNTP浓度以及TaqDNA聚合酶的浓度等对ISSR-PCR扩增的影响;尝试了不同的退火温度、延伸时间和循环次数, 筛选出17个扩增稳定且多态性丰富的ISSR引物;建立了红松ISSR的最佳反应体系,为进行红松种群间遗传分化的研究奠定基础。
【英文摘要】
The study focuses on the influential factors of ISSR experiments used to analyze Koreanpine. Comparative experiments on DNA extraction and PCR amplification among fresh leaves, dryleaves, embryos and endosperms were conducted. The results showed that the four kinds of materi-als were all feasible. The effect of purity and concentration of template, dNTP and TaqDNA poly-merase on amplification was analyzed. Different annealing temperature, extension time and cycletimes were also compared. 17 primers with sta...
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