卷丹百合RAPD-PCR反应程序的优化研究

Optimization of RAPD-PCR Procedure in Lilium lancifolium Thunb.

采用CTAB法提取卷丹百合的基因组DNA。采用正交设计和单因素试验相结合的方法,对卷丹百合RAPD-PCR反应程序中各影响因素进行了优化。结果表明:卷丹百合最佳的RAPD-PCR扩增程序为94℃预变性4 min;94℃变性30 s、39℃退火40 s、72℃延伸1 min,45个循环;72℃延伸10 min。

Genomic DNA of Lilium lancifolium Thunb. was extracted with the method of CTAB. Some influencing factors in the RAPD reaction procedure were studied with orthogonal design and single factor experiment. The results showed that the optimum RAPD-PCR procedure was one cycle of denaturing at 94 ℃ for 4 min, 45 cycles of denaturing at 94 ℃ for 30 s, annealing at 39 ℃ for 40 s, extending at 72 ℃ for 1 min, one cycle of final extension extending at 72 ℃ for 10 min.