从颗粒污泥中提取DNA的几种不同方法比较

Comparison of Different DNA Extraction Methods from Granular Sludge

为了从颗粒污泥中提取出DNA,分别采用CTAB法、TENPC法、Takara试剂盒三种方法提取总DNA,通过提取的核酸总量、纯度、片段分布情况等指标来评价不同的提取方法对颗粒污泥总DNA质量的影响,并考察了三种不同的溶液洗涤样品以及三种不同的细胞破壁方法对提取的DNA质量的影响。结果表明:采用脱腐缓冲液、PBS buffer、EDTA buffer洗涤,液氮研磨破壁后TENPC法提取DNA的效果最好,提取的总量最多,大片段提取效果好,这种方法较适合从颗粒污泥中提取DNA。

Three different DNA extraction methods which were CTAB method, TENPC method and Takara Genomic DNA Extraction Kit and three different washing means and three different lysis ways for granular sludge were com- pared. The effects of them were evaluated based on the yield, purity, molecular size distribution of the recovered DNA. The results showed that using three methods can extract genomie DNA, but the most effective method was TENPC method with grinding in liquid nitrogen after washing samples with humic acids＇ removing buffer, PBS buffer and EDTA buffer. It had the highest total DNA yield and large size fragments, and it was feasible for DNA extraetion from granular sludge.