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 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.
Wu
Y
Y
Department of Medical Genetics, University of Groningen, Antonius Deusinglaan 4, 9713 AW Groningen, The Netherlands.
Stulp
R P
RP
Elfferich
P
P
Osinga
J
J
Buys
C H
CH
Hofstra
R M
RM
D001483:Base Sequence; D003596:Cytosine; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005091:Exons; D005656:Fungal Proteins / Q000235:genetics; D006147:Guanine; D009154:Mutation; D011518:Proto-Oncogene Proteins / Q000235:genetics; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; C098790:MSH6 protein, S cerevisiae; D011518:Proto-Oncogene Proteins; D029701:Saccharomyces cerevisiae Proteins; D006147:Guanine; D003596:Cytosine; D004247:DNA; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila