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以珍贵渐危植物黄山木兰的嫩叶为材料,利用改良CTAB法、SDS法、高盐低p H法和Bio Teke试剂盒法提取基因组DNA,通过超微量紫外分光光度计检测、琼脂糖凝胶电泳、限制性内切酶消化和EST-SSR引物扩增法系统地检测DNA的质量及得率。结果表明,在DNA纯度和得率方面,改良CTAB法提取的DNA结果最佳,Bio Teke试剂盒法提取的DNA纯度较好但得率较低,SDS法和高盐低p H法提取的DNA有明显的蛋白质杂质、多糖及其他次生代谢物和RNA的污染;酶切消化结果中改良CTAB法和Bio Teke试剂盒法表现良好;EST-SSR引物扩增结果中唯有改良CTAB法提取的DNA扩增目的基因条带数目最多且清晰。综合分析,改良CTAB法是最适合黄山木兰叶片基因组DNA提取的方法。
Genomic DNA was extracted from the leaves of Magnolia grandiflora, a plant growing in the most precious and endangered plants, using improved CTAB method, SDS method, high salt and low p H method, and Bio Teke kit method. The supernatant was assayed by ultra-violet spectrophotometer. Sepharose Electrophoresis, restriction endonuclease digestion and EST-SSR primer amplification system for the detection of DNA quality and yield. The results showed that the DNA extracted by modified CTAB method was the best in terms of DNA purity and yield. DNA extracted by Bio Teke kit method had better purity but lower yield. DNA extracted by SDS method and high salt and low p H DNA Obvious protein impurities, polysaccharides and other secondary metabolites and RNA contamination; enzymatic digestion results modified CTAB method and Bio Teke kit method performed well; EST-SSR primer amplification results only improved CTAB method DNA amplified the purpose of the largest number of gene fragments and clear. Comprehensive analysis, modified CTAB method is the most suitable method for the extraction of genomic DNA from Magnolia leaves.