五唇兰基因组DNA提取方法的优化

Optimization of Extraction of Genomic DNA in Phalaenopsis pulcherrima(Orchidaceae)

五唇兰组织内含有大量的酚类、多糖以及此生代谢产物,DNA提取较为困难。为获得高质量的五唇兰基因组DNA,文中采用传统十六烷基三甲基溴化铵(CTAB)法、常规十二烷基硫酸钠(SDS)法和改进的CTAB法对五唇兰基因组DNA的提取进行比较。结果表明:改进的CTAB法比传统CTAB法和常规SDS法都好,可以得到质量较好的DNA,电泳条带清晰,通过紫外分光光度计检测,A260/A280为2.0以上。

Phalaenopsis pulcherrima is rich in polysaccharides,phenol and other secondary metabolites,and it is difficult to obtain high quality of genomic DNA from it.Based on the research results of comparative analysis of several extraction methods,such as traditional Hexadecyltrimethy Ammonium Bromide(CTAB),traditional Sodium Dodecyl Sulfate(SDS) and improved CTAB,for total genomic DNA of P.pulcherrima.The results indicate that improved CTAB DNA extraction produces DNA of much better quality than the others,which has higher density,purity and intact genomic DNA by agarose gel electrophoresis.And by ultraviolet spectrophometry,the ratio of A260/A280 was above 2.0.