Comparison of three DNA extraction methods for polymerase chain reaction (PCR) analysis of bacterial genomic DNA

Abstract

Rapid isolation of high-purity microbial genomic DNA is necessary for genome analysis. In this study, the authors compared a one-hour procedure using a microwave with enzymatic and boiling methods of genomic DNA extraction from Gram-negative and Gram-positive bacteria. High DNA concentration and purity were observed for both MRSA and ESBL strains (80.1 and 91.1 µg/ml; OD260/280, 1.82 and 1.70, respectively) when the extraction protocol included microwave pre-heating. DNA quality was further confirmed by PCR detection of mecA and CTX-M. In conclusion, the microwave-based procedure was rapid, efficient, cost-effective, and applicable for both Gram-positive and Gram-negative bacteria. Key words: DNA, MRSA, ESBL, gene, enzymatic, boiling, microwave.