5种金边瑞香基因组DNA提取方法比较研究

Comparative Study on Five Genomic DNA Extraction Methods of Daphne odora

[目的]研究不同提取方法对金边瑞香基因组DNA提取质量的影响,筛选最佳提取方法。[方法]以金边瑞香叶片为试材,以SDS法、改良CTAB法、SDS-CTAB法、高盐低pH法和尿素法5种方法提取基因组DNA,并从DNA纯度、浓度、酶切电泳结果和ISSR-PCR扩增产物电泳等方面进行比较分析。[结果]高盐低pH法和尿素法提取的金边瑞香基因组DNA富含蛋白质和多糖等杂质,DNA纯度低;SDS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;改良CTAB法提取的基因组DNA纯度较高,但浓度和产率低。SDS-CTAB法提取的基因组DNA纯度、浓度和产率最高,酶切和ISSR-PCR产物电泳检测效果最好。[结论]SDS-CTAB法提取的金边瑞香基因组DNA质量与产量最佳,可以满足后续分子生物学研究的需求。

[Objective]In order to study the effect of different methods on the quality of genomic DNA extraction from Daphne odore,the most suitable extraction method was selected.[Method]Using leaves of Daphne odore as materials,the genomic DNA was extracted by SDS method,modified CTAB method,SDS-CTAB method,high salt low pH method and urea method,which compared and analyzed on the DNA purity,concentration,enzyme digestion electrophoresis results and ISSR-PCR amplification product electrophoresis.[Result]The results showed that the genomic DNA extracted by high salt,low pH method and urea method was rich in impurities such as protein and polysaccharide,that the purity of DNA was low.The genomic DNA extracted by SDS method contains some impurities such as phenols and salts,which the purity of DNA was low.The genomic DNA extracted by modified CTAB method has high purity,but low concentration and yield.The purity,concentration and yield of genomic DNA extracted by SDS-CTAB method were the highest,further more,the effect of enzyme digestion and ISSR-PCR product electrophoresis detection also was the best.[Conclusion]The genomic DNA extracted by SDS-CTAB method has the best quality and yield of Daphne odore,which can meet the needs of subsequent molecular biology research.