Abstract
Denaturing Gradient Gel Electrophoresis (DGGE) is a technique used to separate short- to medium-length DNA fragments based on their melting characteristics. It has been used frequently for identifying single-nucleotide polymorphisms without the need for DNA sequencing and as a molecular fingerprinting method for complex ecosystem communities, in particular in conjunction with amplification of microbial 16S rRNA genes. Here, the principles of DGGE, based on partial DNA strand separation at a given position in a gradient of chemical denaturant, are described, and an example protocol, optimized for fingerprinting of 200–300 bp fragments of bacterial 16S rRNA genes, is given.
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References
Fischer SG, Lerman LS (1979) Length-independent separation of DNA restriction fragments in 2-dimensional gel electrophoresis. Cell 16:191–200
Abrams ES, Murdaugh SE, Lerman LS (1990) Comprehensive detection of single base changes in human genomic DNA using denaturing gradient gel-electrophoresis and a GC clamp. Genomics 7:463–475
Bio-Rad Laboratories (1996) The DCode™ universal mutation detection system (manual). Bio-Rad Laboratories, Hemel Hempstead
Muyzer G, Dewaal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S ribosomal RNA. Appl Environ Microbiol 59:695–700
Felske A, Osborn AM (2005) DNA fingerprinting of microbial communities. In: Osborn AM, Smith CJ (eds) Molecular microbial ecology. Taylor and Francis, Abingdon, pp 65–96
Acknowledgements
This work was supported by Leverhulme Trust a grant no. F/00158/BX, by EPSRC grant no. EP/E030173, and by Darwin Trust Research Fellowship to A.F.
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© 2013 Springer Science+Business Media, New York
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Strathdee, F., Free, A. (2013). Denaturing Gradient Gel Electrophoresis (DGGE). In: Makovets, S. (eds) DNA Electrophoresis. Methods in Molecular Biology, vol 1054. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-565-1_9
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DOI: https://doi.org/10.1007/978-1-62703-565-1_9
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-564-4
Online ISBN: 978-1-62703-565-1
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