A rapid method for genomic DNA extraction from activated sludge treating endosulfan manufacturing wastewater, and its ERIC-PCR fingerprint analysis

A simple and rapid method for cell lyses and direct extraction of intracelluar DNA from activated sludge with minimal shearing was developed. The method was based on extraction with an alkaline buffer, lyses with liquid nitrogen grinding and a high-salt extraction buffer in the presence of sodium dodecyl sulfate (SDS), hexadecyltrimethylammonium bromide (CTAB), and 2-mercaptoethanol. The method only required 4 h, and the yield of 1.041±0.1 mg/g MLSS was achieved. Through 16S rDNA amplification and ERIC-PCR analysis, the method was confirmed could supply high quality DNA fragment for molecular micro-ecology work in activated sludge systems. Besides, the feasibility of ERIC-PCR as a molecular monitoring means for wastewater treatment system was proved.