Abstract
Denaturing gradient gel electrophoresis (DGGE) (Abrams and Stanton 1992; Myers et al. 1987) is a technique suitable for detecting point mutations as well as small deletions and insertions of several base pairs in double stranded DNA. The method is based on the analysis of strand separation (denaturation) of DNA molecules. The process of strand separation by disruption of hydrogen bonds is referred to as “melting” of the DNA, while the “melting point” is defined as the temperature or the denaturant concentration at which half of all possible hydrogen bonds are disrupted. It appears that melting occurs in domains, i. e. stretches of several tens of base pairs melt at the same temperature. Several factors influence the melting of a given DNA sequence: temperature, length, and base composition. GC-rich sequences have higher melting temperatures than AT-rich sequences. Moreover, since hybridization depends on hydrogen-bond formation, the presence of hydrogen-bond disrupting agents (denaturants) in the medium will also affect the melting point. Computer algorithms are available that can predict melting properties of DNA sequences (Lerman and Silver-stein 1987). The resulting melt maps can be used to determine optimal experimental conditions.
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References
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© 1999 Springer-Verlag Berlin Heidelberg
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Netzer, KO. (1999). Denaturing Gradient Gel Electrophoresis. In: Hildebrandt, F., Igarashi, P. (eds) Techniques in Molecular Medicine. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59811-1_6
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DOI: https://doi.org/10.1007/978-3-642-59811-1_6
Publisher Name: Springer, Berlin, Heidelberg
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