摘要
目的:以金钗石斛为材料,建立石斛的RAPD优化条件,应用于石斛相似种的DNA多态性分析。方法:分别设置模板DNA,Mg2+,TaqDNA聚合酶,dNTPs,引物的浓度梯度,优化RAPD反应体系。结果:在25μl总反应体系中,以上反应物的用量分别为100 ng,3.0 mmol/L,1.0 U,0.2 mmol/L,0.4μmol/L时所获得的石斛属DNA指纹图谱谱带型清晰、重复性好。其中用S48扩增的DNA指纹图谱具有更大的相似性,适合于石斛真伪鉴别;而S90扩增的片段具有明显的多态性,适合于石斛遗传分化研究。结论:优化后的RAPD方法可作为研究石斛属植物的遗传多样性及真伪鉴别的有效方法。
Objective： The optimal reaction of RAPD （random amplified polymorphic DNA）of Dendrobium nobile was set up and it was used in determing of plants of Dendrobium DNA polymorphisms. Methods： The different concentration were analyzed from template DNA, Mg^2＋, Taq DNA polymerases, dNTPs,primeron RAPD amplification. Results： In 25 μl RAPD reaction system those concentration were 100 ng,3.0 mmol/L, 1.0 U ,0.2 mmool/L,0.4 μmol/L respectively, and the DNA fringerprintings of Dendrobium were distinct image, reliable stability and good repeatability. The DNA fingerprints amplified with the primer S48 had a similarity, the type of fingerprints still more suited the discrimination of true and fasle Dendrobium; but The fragments of amplifying the genetic DNA of Dendrobium with the primer S90 had more polymorphisms. The type of DNA fingerprints still more suited the study of genetic divergence in Dendrobiurn. Condusion： The optimizing RAPD technique was effective in studying genetic divergence in Dendrobiurn and characterizing species of Dendrobium and its adulterant specise.
出处
《江苏大学学报（医学版）》
CAS
2007年第2期134-137,141,共5页
Journal of Jiangsu University：Medicine Edition
基金
遵义市科技局基金资助项目(200408E-018)
