Method for detecting mutations in DNA, useful especially for diagnosis of familial hypercholesterolemia, comprises specific and then universal amplification

摘要

Method for detecting mutations, insertions, deletions and polymorphisms in DNA, are new. Method for detecting mutations, insertions, deletions and polymorphisms in DNA comprising: (a) performing specific polymerase chain reaction (PCR) using a primer pair that flanks the genomic site being studied, where the 5'-ends of each primer have a universal oligonucleotide sequence (ONS), different for each primer, that is not complementary to the DNA; (b) performing second, universal PCR with a pair of primers labeled, by fluorophores or biotin, that are complementary to ONS; (c) denaturing the PCR products by heating and rapid cooling; (d) separating the PCR products; and (e) determining the migration properties of the products, these properties being dependent on the specific folding and conformation of the individual DNA strands and the specific folding of the DNA sequence. An Independent claim is also included for a kit for the process.