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Extra info for Diagnostic Bacteriology Protocols 2nd Edition (Methods in Molecular Biology Vol 345)
Thus, on-chip PCR allows the single-step amplification and characterization of a DNA sample as a result of separation in liquid- and solid-phase reactions. In contrast to conventional PCR, the reaction is performed directly on the flat surface of a glass slide that holds an array of covalently attached nested primers. The bacterial target DNA is amplified and probed using primers identifying either species-specific sequence regions of ribosomal DNA or unique bacterial target genes, such as virulence or resistance factors.
Chaotropic salts) or polysaccharides that can interact with both the nucleic acids and/or enzymes also may be potent inhibitors (32). There are also compounds that may interfere directly with the polymerase activity or compounds that modify the nucleic acids (29). By adding substances that facilitate the PCR in the presence of inhibitors, or by selectively removing inhibitors from the sample, recent developments have been achieved. The advantage of such approaches is the simplicity and speed (29).
71, 1018–1024. 43. Nogva, H. , Dromtorp, S. , and Rudi, K. (2003) Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR. BioTechniques 34, 804–813. 44. , Nayak, B. , and Kogure, K. (2003) Density-dependent sorting of physiologically different cells of Vibrio parahaemolyticus. Appl. Environ. Microbiol. 69, 3569–3572. 45. Lapizco-Encinas, B. , Simmons, B. , Cummings, E. , and Fintschenko, Y. (2004) Dielectrophoretic concentration and separation of live and dead bacteria in an array of insulators.