Headloop suppression PCR and its application to selective amplification of methylated DNA sequences.

ABSTRACT: Selective amplification in PCR is principally determined by the sequence of the primers and the temperature of the annealing step. We have developed a new PCR technique for distinguishing related sequences in which additional selectivity is dependent on sequences within the amplicon. A 5' extension is included in one (or both) primer(s) that corresponds to sequences within one of the related amplicons. After copying and incorporation into the PCR product this sequence is then able to loop back, anneal to the internal sequences and prime to form a hairpin structure-this structure is then refractory to further amplification. Thus, amplification of sequences containing a perfect match to the 5' extension is suppressed while amplification of sequences containing mismatches or lacking the seque

SUBMITTER: Rand KN 

PROVIDER: S-EPMC1184225 | biostudies-literature | 2005 Aug

REPOSITORIES: biostudies-literature