山兰不同部位DNA提取的比较研究

Comparative Study on Extraction of Genomic DNA from Different Parts of Oreorchis Patens

目的研究珍稀植物山兰的假鳞茎、叶片、花芽中的DNA的质量,为开展分子鉴定等研究提供依据。方法采用CTAB法提取其基因组DNA,用琼脂糖凝胶电泳对其进行质量检测,用分光光度计进行浓度及纯度检测,并分别进行RAPD扩增。结果不同来源的DNA的电泳均有明显条带,表明质量均较好。花芽提取的DNA浓度最高,为356.82μg/μL;叶片次之,为215.43μg/μL;假鳞茎最低,为93.76μg/μL。3个部位提取的纯度相近,不同部位提取的DNA模板对RAPA扩增没有明显影响。结论山兰3个部位提取的质量均较好,可以进行后续的酶切及PCR扩增等实验。从来源上,采用叶片提取DNA最好。

Objective To study the quality of the genome DNA of the pseudo-bulbs, leaves and floral buds of the rare plant Oreorchis patens, and to provide the basis for molecular identification and other research. Method The genome DNA was extracted from three different parts of the plant with CTAB method, examining the quality with agarose gel electrophoresis, measuring the purity and quality with the UV spectrophotometer, and using for RAPD amplification respectively. Result The bands of the DNA from different parts are obvious, which indicates that all are good in quality. The concentration of DNA extracted from its floral buds is the highest（356.82μg/μL） followed by that from leaves（215.43μg/μL）, and that from the pseudobulbs is the lowest（93.76μg/μL）. The purity of DNA from the three parts is similar and the DNA templates from different parts have no obvious effect on RAPD amplification. Conclusion The DNA was extracted from the three parts of the plant all are good in quality and it could be used in the following experiments such as digestion and PCR amplification. And the leaves are the optimal based on the resources.