Patent ID: 8293472

Claim:
A method for detecting a specific mutation in a heterogeneous population of polynucleotides, wherein the specific mutation, if present, is located in a first region of the polynucleotides of the population, the method comprising: (a) amplifying the polynucleotides of the population with a first set of primers to obtain an amplification product that comprises the first region, wherein at least one primer of the first set of primers is a healing primer that specifically hybridizes to a region of the heterogeneous population of polynucleotides that comprises one or more additional mutations, and none of the primers of the first set of primers hybridizes to the first region; and (b) amplifying the amplification product with a second set of primers to detect the specific mutation, wherein the second set of primers comprises a first primer having a nucleotide that is complementary to the first region if the specific mutation is present and the first primer includes at least one non-natural base selected from iso-G or iso-C, and further wherein one or more of the primers of the second set of primers comprises a label.