【摘要】 目的:建立最适的AtSUC9 PCR反应体系。方法:应用降落PCR和正交设计,从Mg2+、dNTPs、引物、甘油4种因素3个水平,对拟南芥AtSUC99 PCR反应体系进行优化。结果:确立了适合拟南芥AtSUC99 PCR反应体系:在20μl反应体系中,含模板1μg,引物(20μmol/L)0.5μl,dNTPs(10mmol/L)0.3μl,10×PCRbuffer2μl,Mg2+(25mmol/L)1.2μl,TaqDNAPolymerase1U,甘油10μl。结论:降落PCR和正交设计是一种有效的植物基因PCR体系优化方法。更多还原

【Abstract】 Objective: A suitable Arabidopsis thaliana AtSUC9 PCR reaction system was established.Methods:In this study,touchdown PCR and orthogonal design was used to optimize PCR reaction system in Arabidopsis thaliana AtSUC9 in four factor (Mg2+,dNTPs,primers,glycerine) at three levels respectively.Results:A suitable PCR reaction system was established,namely 20μl reaction system containing DNA template 1μg,primers (20μmol/L) 0.5μl,dNTPs (10mmol/L) 0.3μl,10×PCR buffer 2μl,Mg2+ (25mmol/L) 1.2μl,Taq DNA Polymerase 1U,glycerine 10μl.Conclusion: Touchdown PCR and orthogonal design proved to be an effiecient PCR reaction system optimum method in plant gene.更多还原