Methods and compositions are provided for detecting a primer extension
product in a reaction mixture. In the subject methods, a primer extension
reaction is conducted in the presence of a polymerase having 3'.fwdarw.5'
exonuclease activity and at least one FET labeled oligonucleotide probe
that includes a 3'.fwdarw.5' exonuclease resistant quencher domain. Also
provided are systems and kits for practicing the subject methods. The
subject invention finds use in a variety of different applications, and
are particularly suited for use in high fidelity PCR based reactions,
including SNP detection applications, allelic variation detection
applications, and the like.