牡丹·芍药DNA提取方法研究

Study on DNA Extraction Methods of Paeonia suffruticosa and Paeonia lactiflora Pall

[目的]筛选一种适合牡丹、芍药基因组DNA的提取方法。[方法]以新鲜的牡丹、芍药叶片为材料,采用CTAB法和SDS法提取基因组DNA,通过紫外光谱分析法、琼脂糖凝胶电泳及PCR扩增等检测方法对牡丹、芍药基因组DNA的提取方法进行筛选。[结果]在牡丹、芍药基因组DNA的提取过程中,叶片内的酚类、色素、丹宁、糖类等物质与DNA结合,影响DNA的提取质量。在这两种DNA提取方法中,SDS法略好于CTAB法。SDS法DNA得率虽低,但其纯度高,去糖去蛋白等杂质比较干净,且在未加RNA酶的情况下,RNA污染较少,在进行PCR扩增时,能得到比较理想的产物。[结论]取材时间是影响DNA提取质量的关键因素之一。在提取牡丹、芍药基因组DNA时应该取完全展开的幼嫩叶片。

[Objective] The research aimed to screen a kind of extraction method for genomic DNA in Poeonia suffruticosa and Paeonia lactiflora Pall. [Method] With fresh leaves of P. suffruticosa and P. lactiflora as materials, CTAB method and SDS method were adopted to extract genomic DNA, And the methods of extracting genomic DNA in P. suffruticosa and P. lactiflora were screened by some detection methods such as UV spectral analysis method, agarose gel electrophoresis and PCR amplification. [Result] During the extraction process of genomic DNA in P. suffruticosa and P. lactiflora, DNA was combined with some substances in the leaves including phenols, pigment, tannin, glucide and so on, which affected DNA extraction quality. Among 2 kinds of DNA extraction methods, SDS method was slightly better than CTAB method. DNA extraction rate of SDS method was low, but it had high purity and could remove impurities such as sugar and protein very clearly. By SDS method, under the condition without adding RNase, RNA population was very little and very ideal products could be obtained in the process of PCR amplification. [Conclusion1 Sampling time was one of key factors that affected the quality of DNA extraction. Entirely expanding tender leaves should be used to extract genomic DNA in P. suffruticosa and P. lactiflora. Keywords