四种海水样品总基因组DNA提取方法的比较研究  

Comparision study on four methods for microbial DNA extraction from seawater

作　　者：尹琦[1,2] 史晓翀[1,2] 董雪[1,2] 富冰冰 刘吉文[1,2] 张晓华[1,2] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003 [2]中国海洋大学海洋生物多样性与进化研究所,山东青岛266003

出　　处：《海洋环境科学》2013年第3期448-450,共3页Marine Environmental Science

基　　金：国家863计划项目(2007AA09Z434);中央高校基本科研业务费(项目号:201122005)

摘　　要：采用4种不同的方法,分别提取了同一来源的50 mL、200 mL和1 000 mL海水样品的基因组DNA,利用紫外分光光度法测定DNA质量和纯度。研究发现:方法 1(使用PIPES缓冲液)和方法 2(使用TENS缓冲液)所提取的DNA,其OD260/OD280和OD260/OD230很低(不足1.4);方法 3(使用STE缓冲液)及其改进后的方法 4(使用STE缓冲液,但利用机械作用取代酶消化作用来破碎细胞)所提取DNA的两个指标都较高(超过1.6),并且方法 4所得DNA的产率是其他3种方法的1.5～2.4倍。另外,检测方法 3和方法 4所提取DNA的PCR扩增效率,结果表明所提取DNA无需纯化就能进行高效PCR扩增反应。综合以上结果,从DNA浓度、纯度及PCR扩增效率可以看出,方法 4是适合于海水样品多样性研究的一种高效DNA提取方法。The uncultultured methods play an important role in studying marine microbial diversity,and the DNA extraction is the key step for these methods.In this study,four different methods were used to extract microbial DNA from three groups of seawater samples（50 mL,200 mL and 1 000 mL）,and UV absorption were used to analyze the quality and quantity of the genomic DNA.The results showed that,DNA purity indexes obtained by method 1（using PIPES buffer） and method 2（using TENS buffer） were less than 1.4,while both the indexes obtained by method 3（using STE buffer） and its improvement-method 4 were over 1.6.The average DNA concentration by method 4（using STE buffer,but used mechanical action instead of enzymic digestion to break cells） was 1.5~2.4 folds of the other three methods.Moreover,the PCR reaction stability of DNA got from method 3 and method 4 was tested,and the results showed that DNA without purification could have stable PCR reactions.We conclude that,considering DNA concentration,purification and PCR efficiency,method 4 was an efficient method to seawater DNA extraction.

关 键 词：海水 微生物 基因组DNA提取 改进方法 

分 类 号：P734[天文地球—海洋化学]