一种快速高效提取革兰氏阳性菌微杆菌基因组DNA的新方法

A Rapid and Efficient Method for Preparing Genomic DNA from Microbacterium sp.

[目的]探索可以快速从革兰氏阳性菌微杆菌中提取高质量基因组总DNA的新方法。[方法]采用3种不同方法提取微杆菌基因组DNA,并进行DNA纯度、完整度鉴定及16SrDNA扩增检测。[结果]紫外吸收结果表明,采用改良方法所获DNAA260/A280大于1.80,A260/A230为2.0,5ml过夜菌液可得6.5μg基因组总DNA。DNA凝胶电泳结果表明,DNA完整度高且无RNA污染。以此方法提取的基因组总DNA样品为模板,可灵敏有效地扩增16SrDNA基因。[结论]该方法用时短,操作简易,纯度好、产率高、成本低,适用于革兰氏阳性菌各相关的分子生物学研究。

[Objective]The aim was to research a new method for preparing genomic DNA from Microbacterium sp. quickly and efficiently. [Method]Three different methods were used to prepared genomic DNA from Microbacterium sp.,and identification of DNA pruity,integrity and 16S rDNA amplification were carried out.[Result]The results of DNA quantity and purity measured by UV absorbance showed that,by the new method,DNA A260/A280 value was greater than 1.8,A260/A230 value was 2.0,and 6.5 μg genomic DNA could obtained from 5 ml overnight broth.The result of agarose gel eletrophoresis indicated high integrity of the genomic DNA and there was no RNA pollution. [Condusion]The DNA prepared by this new method was sufficiently pure for PCR. This method saves time and cost,practices easily as well,and its was applied to all relevant molecular biology related to Gram-positive bacteria.