以PCR为目的的大豆叶片DNA提取方法的比较研究

Comparative Study on Methods of Extracting DNA from Soybean Leaf for PCR

模板DNA的质量直接影响PCR扩增的结果,而不同提取方法及其缓冲液的成份与浓度对提取DNA的质量有重要影响。本文以5个栽培大豆品种的叶片为材料,比较分析了SDS与CTAB两种提取方法以及不同浓度CTAB提取缓冲液对所提取的DNA质量的影响,并通过PCR进行检验。实验结果表明:用1%(W/V)、2%(W/V)浓度的CTAB提取缓冲液和1.25%(W/V)SDS提取缓冲液所提取的大豆叶片DNA的质量较好,均能满足PCR扩增模板的需求,其中以1.25%(W/V)SDS提取得到的大豆叶片DNA质量最好,以其为模板扩增的效果最佳,而4%浓度的CTAB不适宜提取大豆叶片DNA。

Template DNA quality directly affects the effectiveness of PCR amplification. Objective of this study is to explore the effects of extracting-buffer components and their concentrations on PCR-template DNAs from leaf tissues of five soybean cultivars. Genomic DNAs were extracted from soybean leaf tissues of five cultivars by using two kinds of extracting methods, 1.25%(W/V) SDS and CTAB with different buffer concentrations (1%W/V, 2%W/V, 4%W/V). And then the extracted DNA quality was examined through random-primer PCR. The experimental result showed that template DNAs suitable for PCR could be gotten by using 1%, 2% CTAB and 1.25%SDS extraction buffers. Among them, the quality of soybean leaf DNA extracted by using 1.25%(W/V) SDS extraction buffer was the best. Therefore, 1.25%(W/V) SDS was the most suitable method of extracting DNA from soybean leaf tissue for PCR analysis.