SDSC-TAB和高盐沉淀法提取香蕉枯萎病菌基因组DNA的比较

Comparative Study of Genomic DNA from Fusarium oxysporum f. sp. cubense by SDS-CTAB and High-concentration-salt Precipitation Methods

以香蕉枯萎病菌菌株为试验材料 ,采用SDS- CTAB法和高盐沉淀法提纯香蕉枯萎病菌基因组DNA。结果表明 :高盐沉淀法是适合于香蕉枯萎病菌基因组DNA提取的方法。该方法提取的DNAOD2 60 2 80值显示产物纯度较高 ;经琼脂糖凝胶电泳得到一条带型较宽且清晰的DNA谱带 ,DNA浓度较高 ,基本无DNA碎带 ;不用RNase处理 ,已无RNA的干扰 ,无需任何纯化处理即可用于PCR扩增和RAPD分析。同时对DNA提取过程中的细节问题进行了探讨与分析。

SDS-CTAB and High-concentration-salt precipitation methods were used to extract genomic DNA of 10 strains of Fusarium oxysporum f. sp. cubense which were isolated from Hainan banana growing areas. The results show that High-concentration-salt precipitation method was more fit for extracting genomic DNA of Fusarium oxysporum f. sp. cubense than SDS-CTAB method. Genomic DNA extracted with this method had a high purity seen by their OD 260/280 value and had a clear fragment through agarose gel electrophoresis. They do not take RNA without adding RNase and can be directly used as templates for PCR amplification and RAPD analysis without any further purification. Moreover, some detail problems in extraction process of genomic DNA were discussed and analysised.