A method for extraction of high-quality and high-quantity genomic DNA generally applicable to pathogenic bacteria.
In this study, we report a modified procedure for extraction of high-quality genomic DNA that is rapid, simple, biologically nonhazardous, and generally applicable to pathogenic bacteria. Bacterial cells were pretreated with 70% ethanol prior to enzymatic digestion with lysozyme. Exposure of bacterial cells to 70% ethanol sterilized the cultures, making the process biologically safe and increased the susceptibility of the cells to lysozyme-induced lysis. Consistently high yields of genomic DNA (mean average yield, 0.5-2.5 mg/ml) were obtained from 465 isolates representing over 30 clinically important bacterial species. Genomic DNA obtained was determined to be suitable for further analysis, including bacterial fingerprinting techniques like restriction endonuclease analysis, Southern hybridization, and repetitive PCR. Availability of a generally applicable procedure for extraction of high-quality and high-quantity genomic DNA would be immensely beneficial for laboratories engaged in molecular surveillance of nosocomial and community-based outbreaks.
Kalia
A
A
Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110 029, India.
Rattan
A
A
Chopra
P
P
D001433:Bacteriolysis; D001709:Biotechnology / Q000379:methods*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D000431:Ethanol / Q000737:chemistry; D013210:Staphylococcus / Q000737:chemistry* / Q000235:genetics
D004269:DNA, Bacterial; D000431:Ethanol