不同方法提取大豆基因组DNA的效果比较

Study on Extraction Methods of Soybean Genomic DNA

[目的]采用不同方法对大豆基因组DNA进行提取,为大豆基因组DNA的提取提供参考。[方法]采用CTAB法、SDS法和高盐低pH值法对大豆基因组DNA进行提取,对提取的DNA进行光密度、紫外光谱、琼脂糖凝胶电泳和DNA限制性内切酶图谱等的检测。[结果]光密度和光谱检测结果表明,SDS法的提取量和纯度均优于CTAB法和高盐低pH值法;琼脂糖凝胶电泳检测结果表明,SDS法提取的基因组DNA谱带较CTAB法和高盐低pH值法提取的基因DNA谱带粗、亮、重复性好;限制性内切酶检测结果表明,3种方法所得基因组DNA均能被HindⅢ和EcoRⅠ酶切,但SDS法所得基因组DNA的酶切效果最好。[结论]SDS法的提取效果明显优于CTAB法和高盐低pH值法。

[ Objective ] The study was to extract high quality and sufficient yield of genomic DNA of soybean and to provide a reference for the extraction genomic DNA of soybean. [ Method] With soybean seed as material, genomic DNA was extracted by SDS,CTAB and high salt and low pH value method. The genomic DNA was determined through optical density, ultraviolet spectra, agarose gel electrophoresis and the re- striction enzyme digestion of DNA. [ Result ] The results of optical density and spectrum detection showed that , extraction amount and purity with SDS method were better than CTAB method and high salt and low pH value method ; The results of agarose gel eleetrophoresis determina- tion showed that, compared with genome DNA band extracted by high salt and low pH value method, genome DNA band was more crude, brighter and better repeatability extracted by SDS method; The results of restriction endonuclease determination showed that genome DNA ob- talned from three method can be restriction enzyme by HindⅢ and EcoR I , and SDS method was best for the effect of genome DNA of restric- tion enzyme.