Evaluation of DNA extraction methods for the analysis of microbial community in biological activated carbon.
It is difficult to isolate DNA from biological activated carbon (BAC) samples used in water treatment plants, owing to the scarcity of microorganisms in BAC samples. The aim of this study was to identify DNA extraction methods suitable for a long-term, comprehensive ecological analysis of BAC microbial communities. To identify a procedure that can produce high molecular weight DNA, maximizes detectable diversity and is relatively free from contaminants, the microwave extraction method, the cetyltrimethylammonium bromide (CTAB) extraction method, a commercial DNA extraction kit, and the ultrasonic extraction method were used for the extraction of DNA from BAC samples. Spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) analysis were conducted to compare the yield and quality of DNA obtained using these methods. The results showed that the CTAB method produce the highest yield and genetic diversity of DNA from BAC samples, but DNA purity was slightly less than that obtained with the DNA extraction-kit method. This study provides a theoretical basis for establishing and selecting DNA extraction methods for BAC samples.
State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai, PR China.
Lu
L
Zheng
Naiyun
N
Gao
Yang
Y
Deng
D019149:Bioreactors / Q000382:microbiology*; D002606:Charcoal / Q000032:analysis*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D052616:Solid Phase Extraction / Q000295:instrumentation* / Q000379:methods*
D004269:DNA, Bacterial; D002606:Charcoal