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. 1998 Mar 15;26(6):1548–1549. doi: 10.1093/nar/26.6.1548

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

P Guldberg 1, K Grønbak 1, A Aggerholm 1, A Platz 1, P thor Straten 1, V Ahrenkiel 1, P Hokland 1, J Zeuthen 1
PMCID: PMC147432  PMID: 9490806

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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Selected References

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