Temperature-gradient gel electrophoresis as a screening tool for polymorphisms in multigene families

Information about sequence variability between different copies of a multigene family is indispensable for understanding the evolutionary mechanisms acting on multigene families. However, their high copy number has been a major obstacle to systematic analysis. Exemplified by the internal transcribed spacer 1 (ITS1) of the rDNA in Drosophila melanogaster, it is shown how temperature gradient gel electrophoresis (TGGE) can be used to study sequence polymorphisms in a multigene family. Experimental conditions influencing the melting behavior of the ITS1 fragment are discussed as well as discrepancies between observed and calculated melting patterns.