High resolution analysis of point mutations by constant denaturant capillary electrophoresis (CDCE).

Abstract

AbstractIn recent years, the need for techniques capable of detecting and identifying point mutants has increased dramatically in the fields of genomics, cancer research, and molecular diagnostics. The large arsenal of methods for mutation detection ranges from direct sequencing and array hybridization to allele-specific polymerase chain reaction (PCR). CDCE (1) holds a unique position within this list of techniques due to its high flexibility. Having set up a CDCE system, one can use it to solve a wide range of tasks. At one extreme, CDCE is capable of detecting mutants at very low frequencies (as low as 10-6) (2,3), in which it rivals the high sensitivity mutation detection methods like allele specific PCR. At the other extreme, CDCE is capable of identifying mutants with precision comparable to that of direct sequencing (4). In addition, the instrument for CDCE does not need to be dedicated to one purpose. In fact, most capillary DNA electrophoresis systems, such as DNA sequencing capillary instruments, can be adapted for CDCE separations.KeywordsPolymerase Chain ReactionMelting ProfileAllele Specific Polymerase Chain ReactionSeparation MatrixFraction CollectionThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.