Abstract
Standard agarose gel electrophoresis is extensively used to resolve DNA fragments from 0.2 to 40–50 kb. Larger fragments of genomic DNA or whole viral genomes can only effectively be resolved by pulsed-field gel electrophoresis (PFGE), which extends the range of molecular separation from 200 bp to 12 Mb.
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References
Vivanco, A.B., J. Alvarez, I. Laconcha, N. Lopez-Molina, A. Rementeria, and J. Garaizar. 2004. Molecular genotyping methods and computerized analysis for the study of Salmonella enterica. Methods in Molecular Biology 268:49–58.
Swaminathan, B., T.J. Barrett, S.B. Hunter, R.V. Tauxe, and T.F. CDC PulseNet. 2001. PulseNet: the molecular subtyping network for foodborne bacterial disease surveillance, United States. Emerging Infectious Diseases 7:382–389.
Raoult, D., S. Audic, C. Robert, C. Abergel, P. Renesto, H. Ogata, B. La Scola, M. Suzan, and Claverie J.-M. 2004. The 1.2-Megabase Genome Sequence of Mimivirus. Science Fundamentals of Measurement. 306:1344–1350.
Atterbury, R.J., P.L. Connerton, C.E. Dodd, C.E. Rees, and I.F. Connerton. 2003. Isolation and characterization of Campylobacter bacteriophages from retail poultry. Applied & Environmental Microbiology 69: 4511–4518.
Iguchi, A., R. Osawa, J. Kawano, A. Shimizu, J. Terajima, and H. Watanabe. 2003. Effects of lysogeny of Shiga toxin 2-encoding bacteriophages on pulsed-field gel electrophoresis fragment pattern of Escherichia coli K-12. Current Microbiology 46:224–227.
Serwer, P., S.J. Hayes, E.T. Moreno, D. Louie, R.H. Watson, and M. Son. 1993. Pulsed field agarose gel electrophoresis in the study of morphogenesis: packaging of double-stranded DNA in the capsids of bacteriophages. Electrophoresis 14:271–277.
Fuhrman, J.A., J.F. Griffith, and M.S. Schwalbach. 2002. Prokaryotic and viral diversity patterns in marine plankton. Ecological Research 17:183–194.
Diez, B., J. Anton, N. Guixa-Boixereu, C. Pedros-Alio, and F. Rodriguez-Valera. 2000. Pulsed-field gel electrophoresis analysis of virus assemblages present in a hypersaline environment. International Microbiology 3: 159–164.
Breitbart, M., I. Hewson, B. Felts, J.M. Mahaffy, J. Nulton, P. Salamon, and F. Rohwer. 2003. Metagenomic analyses of an uncultured viral community from human feces. Journal of Bacteriology 185:6220–6223.
Edwards, R.A. and F. Rohwer. 2005. Viral metagenomics. Nature Reviews Microbiology 3:504–510.
Carlson, K. 2005. Working with bacteriophages: Common techniques and methodological approaches., In E. Kutter and A. Sulakvelidze (Eds.), Bacteriophages: Biology and Applications. CRC Press, Baco Raton, FL.
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Lingohr, E., Frost, S., Johnson, R.P. (2009). Determination of Bacteriophage Genome Size by Pulsed-Field Gel Electrophoresis. In: Clokie, M.R., Kropinski, A.M. (eds) Bacteriophages. Methods in Molecular Biology™, vol 502. Humana Press. https://doi.org/10.1007/978-1-60327-565-1_3
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DOI: https://doi.org/10.1007/978-1-60327-565-1_3
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