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目的研究菝葜的RAPD-PCR扩增体系最适宜的条件。方法采用CTAB提取方法,从菝葜的嫩叶中提取总基因组DNA,以此为模板,优化菝葜RAPD-PCR的反应条件。结果 最适宜的PCR扩增体系为:反应体积30μL,内含0.20μmol/L引物、200μmol/L dNTP4、0 ng模板DNA、2 UTaqDNA聚合酶、2μmol/L Mg2+。结论通过扩增条件优化实验,建立了重复性好、稳定性高的药用植物菝葜RAPD-PCR扩增体系最适宜的条件,筛选出条带清晰多态性好的14条随机引物,共扩增得到144条电泳带,其中有多态性带62条,检测到了62个多态性位点,多态率为43%。
Objective To study the most suitable conditions of RAPD-PCR amplification system for catfish. Methods The total genomic DNA was extracted from young leaves of Culter by CTAB extraction and used as template to optimize the reaction conditions of RAPD-PCR. Results The most suitable PCR amplification system was as follows: reaction volume 30μL, containing 0.20μmol / L primer, 200μmol / L dNTP4,0 ng template DNA, 2 UTaq DNA polymerase and 2μmol / L Mg2 +. Conclusion The optimum conditions of RAPD-PCR amplification system for medicinal plant RAPD of medicinal plants with good repeatability and stability were established by optimizing the conditions of amplification experiments. Fourteen random primers with good polymorphism were selected and screened out. A total of 144 electrophoretic bands were amplified, of which 62 bands were polymorphic and 62 polymorphic bands were detected. The polymorphism rate was 43%.