CTAB法提取野野村菌基因组DNA

CTAB Method for Genomic DNA Extraction from Nonomuraea

摘要:

针对用常规方法难以提取野野村菌基因组DNA的问题, 通过选用添加甘氨酸的不同培养基和不同培养时间获得的菌丝体, 采用液氮研磨结合CTAB法提取野野村菌DNA, 电泳检测及计算OD260/OD280值。结果表明, 在添加0.3%甘氨酸的麦芽汁-酵母膏(YE)培养基中振荡培养培养3 d的菌丝体适合于DNA提取, 用CTAB法获得的基因组DNA, 长度约为20 kb, 且OD260/OD280在1.8左右, 达到基因组DNA-DNA杂交的要求。

Abstract:

In order to obtain genomic DNA which meets the requirements of DNA-DNA hybridization, CTAB method combined with liquid nitrogen grinding was investigated. Cultures from different media added with glycine were harvested in different time. Quality of extracted genomic DNA was assessed by electrophoresis and OD260/OD280 value. Mycelia of shaking culture in yeast extract & malt extract (YE) medium containing 0.3% glycine for 3 days were subjected for DNA extraction. The genomic DNA obtained was about 20 kb, with the OD260/OD280 value around 1.8.