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Mutation detection by differential primer extension of mutant and wildtype target sequences
Mutation detection by differential primer extension of mutant and wildtype target sequences
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机译:通过突变和野生型靶序列的差异引物延伸进行突变检测
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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.
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