Detection of mutations in GC-rich DNA by bisulphite denaturing gradient gel electrophoresis. |
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| Authors: | P Guldberg, K Gr nbak, A Aggerholm, A Platz, P thor Straten, V Ahrenkiel, P Hokland, J Zeuthen |
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| Affiliation: | P Guldberg, K Grønbak, A Aggerholm, A Platz, P thor Straten, V Ahrenkiel, P Hokland, and J Zeuthen |
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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. |
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