几种水稻田土壤微生物总DNA提取方法的比较  

Comparison of Methods of DNA Extraction from Paddy Soil

作　　者：顾华杰[1] 李玉祥[1] 赵明文[1] 潘根兴[2] 

机构地区：[1]南京农业大学农业部农业环境微生物工程重点开放实验室,江苏南京210095 [2]南京农业大学农业资源与生态环境研究所,江苏南京210095

出　　处：《江苏大学学报（医学版）》2005年第4期300-305,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(40231016)

摘　　要：目的:使人们在提取土壤总DNA时,可以根据不同的要求选用不同的方法。方法:选用了5种现在较常用的土壤微生物总DNA提取方法从水稻田土壤中提取总DNA,并对5种方法提取的DNA的质量和数量进行比较评价。结果:5种方法都可以从土壤中提取到长度大于48 kb的DNA片段,但不同方法提取的DNA的产量存在明显差异。土壤总DNA都不需要纯化就可以用于PCR扩增,使用细菌16S rDNA基因的引物可以扩增得到相应的片段。结论:Bourrain法和M artin-Laurent法可以较快地提取出DNA,M artin-Laurent法提取的DNA的量最多,Tied je法和Jans-sen法提取的DNA片段最大,Janssen法提取的DNA纯度最高,Reddy法提取的DNA的完整性和PCR扩增效果最好。Objective： To make it possible that when DNA is extracted from soil, different methods could be chosen according to different demands, Methods： In this experiment, five different methods commonly used to extract DNA from soil were chosen to extract DNA from paddy soil. Then the quality and quantity of DNA were compared to estimate the five methods, Results ： DNA extracted by five methods were all greater than 48kb in size, but the yields were different. Subsequently, they were directly used as tem- plates for PCR amplification without purification, using primers of the bacterial 16S rDNA gene. Conclusion： Bourrain method and Martin-Laurent method can generate DNA quicker than other methods. MartinLaurent method can generate the largest amount of DNA. Tiedje method and Janssen method can generate the largest DNA fragments in size. Janssen method can generate the purest DNA. Reddy method can generate DNA in highest integrity which was the most suitable for PCR amplification.

关 键 词：水稻田土壤 微生物 方法比较 DNA提 

分 类 号：Q939.96[生物学—微生物学]