title: Mutagenically separated PCR (MS-PCR): a highly specific one step procedure for easy mutation detection.
abstract: With increasing knowledge about the causal role of genetic defects in clinical diseases the necessity is apparent to have procedures for rapid diagnosis of point mutations. We developed a PCR-based technique, whereby both normal and mutant alleles can be amplified in the same reaction tube, using different length allele-specific primers. Furthermore the allele-specific primers introduce additional deliberate differences into the allelic PCR-products that drastically reduce crossreactions in subsequent cycles. This mutagenesis separates the amplification reactions of the alleles performed in the same tube. Subsequent identification of the PCR-products is done by gel electrophoresis and shows at least one of the two allelic products. Therefore, in addition to simple handling, MS-PCR provides a within-assay quality control for the exclusion of false negative results. The feasibility of this technique has been tested using six different mutations. The high sensitivity of MS-PCR also allows screening for mutation carriers in pooled DNA samples.
authors.affiliation: Institut für Arterioskleroseforschung, Münster, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Rust
authors.forename: H
authors.initials: H
authors.lastname: Funke
authors.forename: G
authors.initials: G
authors.lastname: Assmann
mesh_terms: D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D004247:DNA; D006801:Humans; D006937:Hypercholesterolemia / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D001055:Apolipoproteins B; D004247:DNA