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

Authors: Guldberg, Per¹; Grønbæk, Kirsten¹; Aggerholm, Anni²; Platz, Anton³; Straten, Per thor¹; Ahrenkiel, Vibeke¹; Hokland, Peter²; Zeuthen, Jesper¹

Source: Nucleic Acids Research, Volume 26, Number 6, 15 March 1998 , pp. 1548-1549(2)

Publisher: Oxford University Press

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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.

Document Type: Research article

Affiliations: 1: Department of Tumour Cell Biology, Institute of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, DK-2100 Copenhagen, Denmark, 2: Department of Haematology, Aarhus University Hospital, Aarhus, Denmark and 3: Department of Oncology, Karolinska Hospital, Stockholm,Sweden

Publication date: 1998-03-15

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Nucleic Acids Research (NAR) is a fully Open Access journal, providing rapid publication of leading edge research into the nucleic acids under the following categories: chemistry, computational biology, genomics, molecular biology, nucleic acid enzymes, RNA and structural biology. There is a Survey and Summary section, and methods papers are published
in NAR Methods Online. Each year the first issue is devoted to biological databases, and a later issue to relevant web-based software resources.