Serial coupling of restriction cleavage and extension for nucleic acid amplification

摘要: A novel method of pyrophosphorolysis activated polymerization (PAP) has been developed. In PAP, and by DNA polymerase are coupled serially for each amplification using an activatable oligonucleotide P* that a non-extendable 3′-deoxynucleotide at its 3′ terminus. PAP can be applied exponential or linear amplification. to rare allele in admixture with one more wild type alleles is exact match end the but mismatch near terminus allele. inhibited specific subsequence as far 16 nucleotides away from greatly increase specificity detection extremely mutant presence Specificity results both since significant nonspecific requires combination misincorporation polymerase, event. Using genetically engineered polymerases improves efficiency PAP.