Example of allele-specific real-time PCR amplification curves of cloned wild type and mutant TGT plasmid DNAs at different frequencies (raw data).

<p>The specificity and accuracy of the assay was determined by running allele-specific real-time PCR of mixtures of mutant and wild type DNA standards ranging from 0.01 to 100% and amplified with the mutant specific primers in the background of wild type sequence. Amplification of the total population results always in the same Ct values, regardless of the amount of mutant DNAs present in the reaction. WT, wild type.