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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-62703-565-1_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-564-4
Online ISBN: 978-1-62703-565-1
eBook Packages: Springer Protocols