title: Detection of mutations in GC-rich DNA by bisulphite denaturing gradient gel electrophoresis.
abstract: Denaturing gradient gel electrophoresis (DGGE) in combination with PCR and 'GC-clamping' has proven highly efficient as a method for detection of DNA sequence differences. Due to strand dissociation phenomena, however, its use has been limited to the analysis of sequences with a relatively low content of GC pairs. This paper describes how treatment of template DNA with sodium bisulphite drastically lowers the melting temperature of very GC-rich sequences and renders them amenable to DGGE analysis. We demonstrate the use of bisulphite DGGE for rapid and efficient detection of mutations in the p16(INK4/CDKN2) tumour suppressor gene.
authors.affiliation: Department of Tumour Cell Biology, Institute of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, DK-2100 Copenhagen, Denmark. perg@bio.cancer.dk
authors.forename: P
authors.initials: P
authors.lastname: Guldberg
authors.forename: K
authors.initials: K
authors.lastname: Grønbak
authors.forename: A
authors.initials: A
authors.lastname: Aggerholm
authors.forename: A
authors.initials: A
authors.lastname: Platz
authors.forename: P
authors.initials: P
authors.lastname: thor Straten
authors.forename: V
authors.initials: V
authors.lastname: Ahrenkiel
authors.forename: P
authors.initials: P
authors.lastname: Hokland
authors.forename: J
authors.initials: J
authors.lastname: Zeuthen
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004586:Electrophoresis / Q000379:methods*; D005069:Evaluation Studies as Topic; D019942:Genes, p16; D006801:Humans; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D013447:Sulfites
chemical_list: D017931:DNA Primers; D013447:Sulfites; D004247:DNA; C009279:sodium bisulfite