优化麝香百合RAPD反应体系的研究

目的]建立适合百合种质的RAPD优化体系。方法]从麝香百合中提取基因组DNA,进行RAPD反应。通过正交试验设计,优化麝香百合的RAPD反应条件和反应体系。结果]优化的RAPD反应条件为:DNA 50 ng,Primer 0.3μmol/L,Taq酶1.0 U,dNTPs 200μmol/L,Mg2+3.0 mmol/L。在麝香百合的RAPD扩增程序中,各因素显著性程度依次为:循环次数>延伸时间>复性温度>复性时间。麝香百合的最佳RAPD反应程序为:37℃复性50 s,延伸60 s,循环35次。结论]该研究建立了一套稳定的RAPD反应体系,具有较好的扩增效果。

Optimization of RAPD Reaction System for Lilium formolongi

Objective] The aim of the research was to set up a kind of RAPD optimized system for lily germplasm.Method] Genomic DNA was extracted from Lilium formolongi to make RAPD reaction.Through the orthogonal experimental design,RAPD reaction conditions and reaction system for L.formolongi were optimized.Result] The optimized conditions of RAPD reaction were as follows: DNA 50 ng,Primer 0.3 μmol/L,Taq 1.0 U,d NTPs 200 μmol/L and Mg2+ 3.0 mmol/L. In RAPD amplification program of L.formolongi,the significance of each factor in order was as cycle number>elongation time>annealing temperature>annealing time.The optimum RAPD reaction program for L.formolongi was as follows: annealing for 50 s at 37 ℃,elongating for 60 s,recycling for 35 times.Conclusion] A set of stable RAPD reaction system was set up in this research and it had better amplification effects.