Journal article
Autoclave method for rapid preparation of bacterial PCR-template DNA
Journal of microbiological methods, Vol.56(2), pp.143-149
02/01/2004
PMID: 14744443
Web of Science ID: WOS:000189084000001
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Abstract
An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility. Bacteria are lysed by rapid heating and depressurization in an autoclave. The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration. This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components. Bacteria examined were
Enterococcus and
Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat. DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis. The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost.
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Details
- Title
- Autoclave method for rapid preparation of bacterial PCR-template DNA
- Publication Details
- Journal of microbiological methods, Vol.56(2), pp.143-149
- Resource Type
- Journal article
- Publisher
- Elsevier B.V
- Copyright
- © 2003 Elsevier B.V. All rights reserved.
- Identifiers
- WOS:000189084000001; 99380468035006600
- Academic Unit
- Hal Marcus College of Science and Engineering ; Center for Environmental Diagnostics and Bioremediation
- Language
- English