Detection of mutations in GC-rich DNA by bisulphite denaturing gradient gel electrophoresis | Zendy

Per Guldberg | Zendy; Kirsten Grønbæk | Zendy; Anni Aggerholm | Zendy; Anton Platz | Zendy; Per thor Straten | Zendy; Vibeke Ahrenkiel | Zendy; Peter Hokland | Zendy; Jesper Zeuthen | Zendy

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Detection of mutations in GC-rich DNA by bisulphite denaturing gradient gel electrophoresis

Author(s) -

Per Guldberg,

Kirsten Grønbæk,

Anni Aggerholm,

Anton Platz,

Per thor Straten,

Vibeke Ahrenkiel,

Peter Hokland,

Jesper Zeuthen

Publication year - 1998

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/26.6.1548

Subject(s) - biology , temperature gradient gel electrophoresis , gel electrophoresis , dna , microbiology and biotechnology , gene , dna sequencing , genetics , 16s ribosomal rna

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.

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