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Stephan Rust, Harald Funke, Gerd Assmann, Mutagenically separated PCR (MS-PCR): a highly specific one step procedure for easy mutation detection, Nucleic Acids Research, Volume 21, Issue 16, 11 August 1993, Pages 3623–3629, https://doi.org/10.1093/nar/21.16.3623
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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-speclflc primers. Furthermore the allelespeclfic primers Introduce additional deliberate differences Into the allellc PCR-products that drastically reduce crossreactions In subsequent cycles. This mutagenesls 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, MSPCR provides a withln-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.
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