Mutation analysis of TP53 exons 5-8 by automated constant denaturant capillary electrophoresis

DNA fragments melt characteristically according to their nucleotide sequenc e and length, when exposed to denaturants such as temperature, urea or form amide. Small differences within a defined sequence, like a base mutation, w ill result in a slightly different melting behavior of the aberrant DNA fra gment compared to that of the wild type sequence. This feature has previous ly been exploited for mutation detection by constant denaturant capillary e lectrophoresis (CDCE). In this report, we describe an automated approach (A CDCE) using a commercially available apparatus (ABI 310 Genetic Analyzer) t o analyze mutations in exons 5-8 of TP53. The running conditions were deter mined by temperature titration of the fragments on the apparatus, and an op erating sensitivity showed that 0.1% mutated alleles could be detected agai nst a background of wild-type alleles. Up to 48 samples can be analyzed by ACDCE without any need for operator intervention. The apparatus is commerci ally available, and there is no need for instrument modification. To our kn owledge this is the first report on the analysis of TP53 exons 5-8 by ACDCE . Copyright (C) 2001 S. Karger AG, Basel.