乳品废水活性污泥总DNA提取方法的比较

为了从乳品废水活性污泥中快速提取总DNA,本文系统对SDS高盐缓冲液抽提法、冻融+SDS高盐缓冲液抽提法、溶菌酶+SDS抽提法与冻融+溶菌酶+SDS抽提法等4种抽提活性污泥总DNA方法做了比较研究,并对提取的DNA浓度以及通过PCR扩增技术对DNA的质量进行了检测。结果表明:4种方法均可从乳品废水活性污泥中提取出DNA4,种方法提取的DNA总量与纯度存在差异,其中采用冻融+SDS高盐缓冲液抽提活性污泥总DNA效果最好,以该方法提取的总DNA为模板,PCR扩增16 SrDNA,可扩增出分子量为1.5 Kb左右的DNA产物。本文为研究乳品废水活性污泥总DNA的提取提供了一种简便、快速的方法。

Comparison of Different DNA Extraction Methods from Dairy Wastewater Activated Sludge

In order extract total DNA from dairy wastewater activated sludge rapidly and effectively,four DNA extraction methods such as SDS-high salinity buffer fluid lysing method,SDS-high salinity buffer fluid-freezing thaw lysing method,Lysozyme-SDS-freezing thaw lysing method and Lysozyme-SDS lysing method were compared and investigated.Concentration of the extracted DNA and DNA quality were tested by PCR amplification.The results showed that all these methods can extract certain DNA fragment from activated sludge but with different DNA amount and purity.SDS-high salinity buffer fluid-freezing thaw lysing method obtained the highest total DNA yield.With the total DNA extracted by this method as template,about 1.5 Kb of DNA can be obtained through PCR amplification of 16S rDNA.This study provides a simple and rapid method for extracting total DNA from dairy wastewater activated sludge.