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PNA Clamp Technique for Detecting a Ki-ras2 Mutation Using the LightCycler Instrument
Literature indicates that point mutations in codon 12 of the Ki-ras2 gene are associated with colon cancer. The detection of a point mutation in the high background of wild-type cells is very difficult, which represents a problem for many research projects focused on processes that take place during cancerogenesis. Therefore, a quick and easy, yet reliable method of detecting single point mutations is preferred. This article shows an application example for Peptide Nucleic Acid (PNA) oligomers in combination with Hybridization Probes on the LightCycler Instrument. Here, PNAs suppress the codon 12 wild-type PCR product, as they bind to nucleic acids with higher stringency, compared to deoxyribonucleotides, and do not serve as primers for Taq polymerase. Due to these properties, PNAs can be used to detect single point mutations in research samples in a high background of wild-type sequences.
Roche Life Science | High Pure
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Roche Life Science | Gene Expression
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