不同环境中土壤微生物总DNA的提取与纯化

Extraction and purification of total DNA of soil microorganisms in different environments

采用SDS高盐缓冲液抽提法提取了林地、厂区、菜地3种不同环境中土壤微生物总DNA，结果表明，该方法可以从土壤微生物中提取到长度大于23．1kb的DNA；不同环境中提取的DNA产量和纯度都存在一定的差异，每克干土的DNA提取量介于53．55～579．93μg之间，其中以处于原始状态的丛林提取量最高。粗提的土壤微生物DNA采用酚氯仿和柱式腐殖酸去除剂进行纯化，纯化后的土壤微生物DNA可以用于PCR扩增，使用细菌16SrDNA基因的引物可以扩增到相应的片段。

In this experiment, the total DNA of soil microorganisms from forest land, factory area and vegetable plot were extracted by high salt buffer solution of SDS extraction method. The results showed that the extracted DNA length of soil microorganisms was over 23.1 kb by this method. The DNA amount and purity of different soil microorganisms were different, the extracted DNA amount was 53.55-579.93 μg per gram dry soil,and the extracted DNA amount of soil microorganisms in original forest was the highest. The crude extracted DNA of soil microorganisms was purified by phenolchloroform and column humic acid, then purified DNA could be used as templates for PCR amplification, and corresponding fragments could be amplified by using primers of bacterial 16S rDNA gene.