锐尖山香圆基因组DNA提取方法比较研究

Comparison of different extraction methods for Turpinia arguta genomic DNA

为了研究不同方法对锐尖山香圆基因组DNA提取质量的影响,以选择出最适宜的提取方法。以锐尖山香圆叶片为试验材料,采用CTAB法、SDS法、高盐低pH值法和尿素法提取其基因组DNA,并对提取的基因组DNA OD_(260/280)值、OD_(260/230)值、DNA浓度、DNA产率、琼脂糖凝胶电泳、酶切验证和ISSR-PCR扩增产物进行比较分析。结果表明,应用高盐低pH值法和尿素法提取的基因组DNA含有蛋白质和多糖等杂质较多,DNA纯度较低;CTAB法提取的基因组DNA含有一些酚类物质和盐等杂质,DNA纯度较低;SDS法提取的基因组DNA纯度、浓度和产率最高,酶切和ISSR-PCR检测效果最好。SDS法提取的锐尖山香圆基因组DNA质量和产量最好,可用于后续相关分子生物学研究。

In order to study the effect of different methods on the quality of genomic DNA extracted from Turpinia arguta,the most suitable extraction method was selected.The genomic DNA was extracted by using CTAB method,SDS method,high salt low pH method and urea method.The genomic DNA OD_(260/280) value,OD_(260/230) value,DNA concentration,DNA yield,agarose gel electrophoresis,enzyme digestion test and ISSR-PCR amplification products were compared and analyzed.The genomic DNA extracted by high salt low pH method and urea method contained much impurities such as protein and polysaccharide,and the DNA purity was lower;The genomic DNA extracted by CTAB method contains some impurities such as phenols and salts,and the DNA purity was low;The purity,concentration and yield of genomic DNA extracted by SDS method are the highest,and the results of enzyme digestion and ISSR-PCR are the best.The quality and yield of genomic DNA extracted by SDS method are the best,which can be used for subsequent related molecular biology research.