MUTATION DETECTION BY DIFFERENTIAL PRIMER EXTENSION OF MUTANT AND WILDTYPE TARGET SEQUENCES

摘要

The present invention provides methods for simultaneously interrogating two or more related polynucleotides. One embodiment of the present invention concerns a method for simultaneously analyzing a genetic mutation and a corresponding wild-type sequence within a sample. This method comprises: a) hybridizing a primer to a nucleic acid suspected of containing a genetic mutation, wherein the primer is hybridized 3' to the suspected mutation; b) extending the primer in the presence of a mixture of at least one deoxynucleoside triphosphate and at least one chain terminating dideoxynucleoside triphosphate selected such that the wild-type extension product and the mutant-DNA derived extension product have a total number of nucleotides that differ from one another and from the primer; c) separating the primer, the mutant DNA-derived extension product and the wild-type extension product on the basis of their size; and d) identifying the mutant DNA-derived extension product and the wild-type extension product.