Improved mutation detection in GC-rich DNA fragments by combined DGGE and CDGE

Denaturing gradient gel electrophoresis (DGGE) has proven to be a powerful pre-screening method for the detection of DNA variants. If such variants occur, however, in DNA fragments that are very rich in G and C, they may escape detection. To overcome this limitation, we tested a novel gel system whi...

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Bibliographic Details
Published in:Nucleic acids research 1999-08, Vol.27 (15), p.e9-9
Main Authors: Wu, Y, Stulp, R P, Elfferich, P, Osinga, J, Buys, C H, Hofstra, R M
Format: Article
Language:English
Subjects:
Base Sequence
Cytosine
DNA - chemistry
DNA - genetics
DNA-Binding Proteins
Drosophila Proteins
Electrophoresis, Polyacrylamide Gel - methods
Exons
Fungal Proteins - genetics
Guanine
Mutation
Proto-Oncogene Proteins - genetics
Proto-Oncogene Proteins c-ret
Receptor Protein-Tyrosine Kinases - genetics
Saccharomyces cerevisiae Proteins
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Summary:Denaturing gradient gel electrophoresis (DGGE) has proven to be a powerful pre-screening method for the detection of DNA variants. If such variants occur, however, in DNA fragments that are very rich in G and C, they may escape detection. To overcome this limitation, we tested a novel gel system which combines DGGE and constant denaturant gel electrophoresis (CDGE), as it might have the advantages of both methods. Indeed, this combination had the advantages of both methods, good separation of hetero-duplex molecules and prevention of total strand dissociation, and it proved successful in the detection of DNA variants in several GC-rich fragments.
ISSN:1362-4962
0305-1048
1362-4962
DOI:10.1093/nar/27.15.e9