Method for detection of mutant alleles combining real time PCR and REMS-PCR

摘要

The invention provides an amplification system for the simultaneous detection of mutant alleles, and identification of the specific mutated sequence. A sample is enriched and simultaneously genotyped by the presence of peptide nucleic acids (PNAs) probes in an homogeneous single tube amplification reaction, by detecting the cleavage of a fluorophore quencher from the 5′-end of PCR products and, concurrently, selecting DNA that includes specific mutations relative to wild type, by employing a thermostable endonuclease that will only cleave an amplicon formed on a mutation bearing template strand. Oligonucleotides and kits for conducting the amplification system are also provided.