美味猕猴桃基因组DNA的高效提取

Efficient Methods for Genomic DNA Extraction from Actinidia Deliciosa

为了从富含多糖和多酚等次生代谢物质的猕猴桃叶片及野外采集保存的叶样中分离出高质量的基因组DNA,以美味猕猴桃幼叶为试材,对4种不同的DNA提取方法与3种不同的样品保存处理的DNA提取效果进行了试验研究,并首次采用CTAB区室法提取猕猴桃基因组DNA。结果表明,CTAB区室法与硅胶脱水干样保存处理所提取的DNA效果最佳,其OD260/OD280值为1.8以上,能完全被EcoRI酶切消化,1%琼脂糖凝胶电泳谱带明亮清晰,并能获得条带清晰、多态性好的PCR扩增图谱。

In order to isolate high quality DNA from Actinidia leaves enriching polysaccharides and polyphenols and preserved from field samples, the effects of DNA extraction were studied with four different methods including CTAB regular method, CTAB subarea method, SDS regular method, SDS subarea method and three different conditions of storing sample (frozen at-20℃ and-70℃, dried with silica gel). Moreover, the CTAB subarea method was used for the first time for genomic DNA isolation from Actinidia .The results showed that the CTAB subarea method and dried with silica gel was optimal, the value of OD260/OD280 was over 1.8,the DNAs were completely digested by restriction endonuclease and the extracted DNA by this method was appeared as a clear band when electrophoresed in 1% agrarose gel, and produced clear polymorphic patterns amplified by PCR .