Patent ID: 8076082

Claim:
A method for surveying the status of multiple mutation markers in a large background of wild-type DNA, the method comprising: providing a sample including mutant DNA and wild-type DNA, the mutant DNA including mutations; amplifying the sample by a first PCR to generate DNA fragments containing the mutation sites; wherein the first PCR is multiplexed PCR utilizing two or more pairs of PCR primers, wherein each pair of primers includes a forward primer and a reverse primer, wherein each pair of primers is adapted to amplify a sequence containing one or more of the mutation sites and wherein each primer includes both a target-specific part and a universal tail part, wherein the target-specific part flanks the sequence to allow for amplification of the sequence and the sequence of the universal tail part is the same for all forward and reverse primers, wherein the universal tail cannot bind to human genomic sequence; enriching the mutant DNA fragments containing the mutations from the amplicons of the multiplexed PCR simultaneously by performing one or multiple mutant-specific enrichment cycles including competing mutation-specific hybridization and extraction to thereby form an enriched system; amplifying the enriched system by a second PCR to generate sufficient amounts of mutant DNA for detection; and simultaneously surveying the status of the target mutation sites.