Chembiochem. 2015 Jan 19;16(2):209-13. doi: 10.1002/cbic.201402640. Epub 2014 Dec 22.
Simultaneous genotyping of multiple somatic mutations by using a clamping PNA and PNA detection probes.
Kim YT1, Kim JW, Kim SK, Joe GH, Hong IS.
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Abstract
It has been very difficult to detect and quantify multiple somatic mutations simultaneously in single-tube qPCR. Here, a novel method for simultaneous detection of multiple mutations and a melting curve analysis was developed by using clamping PNA and detection PNA probes. Each PNA probe was designed to have a specific melting temperature by the introduction of γ-PNA monomer. This technique was successfully applied to the detection of six genotypes in two different mutations of K-RAS at the same time. Such simultaneous analysis of an amplified curve and a melting curve in qPCR can be widely used for the early diagnosis of cancer and determining the prognosis of drug treatments.
© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
KEYWORDS:
cancer; gamma-PNA; multiple genotyping; peptide nucleic acid; quantitative PCR; somatic mutations
