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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.

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Product Information (3)

Roche Life Science | Gene Expression

Perform a complete gene expression workflow with quality reagents, instruments, and consumables from Roche.

Roche Life Science | High Pure

Get flexible isolation of highly pure nucleic acid from a variety of sample materials with Roche High Pure Kits.

Roche Life Science | FastStart

Consistently achieve sensitive, specific, and reproducible hot start PCR results with the FastStart qPCR product line from Roche.

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