Abstract
MeltMADGE reconfigures the mutation scanning process of denaturing gradient gel electrophoresis (DGGE) so that the independent variable is time rather than space and the dependent (denaturing) variable is temperature rather than concentration of chemical denaturant. Use of a thermal ramp enables use of a homogeneous gel and therefore of high density arrays of wells such as those of microplate array diagonal gel electrophoresis (MADGE). In this configuration, electrophoresis of products on 10-12 96-well meltMADGE gels can be conducted in a 1-2 litre tank in a 1-2hour run, enabling the scanning of a target amplicon in over 1,000 subjects simultaneously. Gels are read by imaging fluorescence of UV-excited ethidium bromide, giving a simple, economical system for identifying rarer sequence variants in target genes which is suitable for large-scale case-control or population studies and other comparable applications. Different amplicons with similar melting characteristics can also be combined into the same run.