大豆基因组DNA提取纯化方法研究

Study on Extraction and Purification Methods of Soybean Genomic DNA

[目的]通过逐步富集的方法，提高大豆DNA的纯度和质量，为其他动植物基因组DNA的提取纯化提供参考。[方法]以梅桥大豆为试验材料，在CTAB法提取大豆基因组DNA的基础上，对大豆基因组DNA进行了纯化，对纯化后的DNA进行了光密度、紫外光谱、琼脂糖电泳和RAPD—PCR等的检测。[结果]光密度和紫外光谱检测表明A260／A280介于1．678～1．722，平均为1.697，A260／A280介于1．733—2．022，平均为1.844；琼脂糖电泳检测证实DNA分子量较大、完整性好、RNA残留少；RAPD—PCR检测得到了谱带清晰、多态性丰富的电泳谱带。DNA的得率为66．969μg／g。该试验的研究方法具有DNA质量高、操作简便、试验条件要求不高的特点。[结论]该方法可以应用于其它动植物的DNA提取纯化。

[ Objective] The study was to improve the purity and quality of soybean genomic DNA with gradually enriching method and to provide a reference for the extraction and purification of other animal and plant genomic DNA. [ Method] With Meiqiao soybean as material, soybean genomic DNA were purified on the basis of extracting soybean genome DNA by CTAB method. The purified DNA was determined through optical density, ultraviolet spectra, agarose gel electrophoresis and RAPD-PCR, etc. [ Result] The detection by optical density, ultraviolet spectra showed that A260/A280 was from 1. 678 to 1. 722 with an average of 1. 697, and the A260/A280 was from 1. 733 to 2. 022 with a mean value of 1. 844. The agarose gel electrophoresis showed the soybean genomic DNA had greater molecular weight, well integrity and little RNA residue. The clear and rich polymorphism bands were obtained through the detection of RAPD-PCR reaction. The yield of soybean genomic DNA was 66. 969 μg/g. The method studied by this experiment could get the soybean genomic DNA with high quality and had the characteristic of simple operation and not high requirement of experimental condition. [ Conclusion ] The method could be applied to purify DNA of other animal and plants.