大豆不同生长时期基因组DNA提取方法的优化

Extraction Method Optimization of Soybean DNA from Different Growth Periods

以探索大豆嫩叶期基因组提取方法为基础,探索从富含酚类、多糖等代谢物质的大豆开花期叶片中获得高质量DNA的提取方法。采用不同的方法提取5种大豆品种在不同生长时期的叶片基因组,并通过对提取的DNA的含量测定、琼脂糖凝胶电泳分析和内参基因的PCR扩增,对不同方法所提取的DNA质量进行综合评价。在嫩叶期,利用前人改进的CTAB法能够分离出纯度高、质量好的DNA,而在开花期,采用本试验新改良的CTAB法可以更有效去除多糖、酚类等物质得到完整性和纯度较好的大豆基因组。前人改进的CTAB法和本试验摸索的新改良CTAB法分别适用于嫩叶期、开花期大豆基因组DNA提取,为大豆的分子生物学研究奠定了基础。

varieties as experimental materials, different genomic DNA extraction methods were conducted and the concentration of genome DNA was detected. Comprehensive evaluation of the DNA quality from different extraction methods was conducted by agarose electrophoresis and PCR amplification of internal control gene. The results indicated that, in fresh leaves period, DNA extracted with the modified CTAB method had higher quality and purity. However, in flowering stage, most ,ff polysaccharides and polyphenolic compounds could be removed more efficiently from the DNA extracted by new modified CTAB method. The results indicated that the new method was suitable for extracting high qualified DNA. The researches indicated that the predecessors＇ modified CTAB method and the new modified CTAB method conducted in this study were applicable to soybean genome DNA extraction in fresh leaves period and flowering stage respectively. This study laid a foundation of soybean molecular bioresearch.