Optimization of profiles of PCR-denaturing gradient gel electrophoresis for anaerobic activated sludge

The optimization of PCR-DGGE atlas for special sludge has attracted the emphasis of the investigation of microbial molecular ecology. In the research of denitrification inhibiting sulfate-reducing bacteria, DNA extracting method of anaerobic sulfate reducing sludge and denitrifying sludge was studied. By amplifying 16S rDNA sequence with universal primer 958F/1401R, the influences of different annealing temperatures on DGGE fingerprint were investigated, and the DGGE adas was optimized by time travel. The results indicate that it is beneficial to the DNA extraction from sulfate reducing sludge by PBS washing sludge and sonic oscillation; the amplified segment is about 500 bp with the primer 958F/1401R, and the influences of different annealing temperatures on DGGE fingerprint are not obvious; the isolating effects of different electrophoresis stages for samples can be reappeared very well by time travel, and the best denaturing gradient ranks from 40% to 60%, the best electrophoresis time is 5 h under 130 V voltage.