Somatic mutations – detecting less than 0.05% KRAS and BRAF mutation in a background of wildtype DNA

Christina Andersen1, Tine Y. Wollff2, Alice Riva3, Milo Frattini3, Ulf Bech Christensen1, Majbritt H. Kyneb2
1PentaBase ApS, 2Danish technological institute, Life Science department, 3Laboratory of Molecular Pathology, Institute of Pathology Via in Selva

By using modified oligonucleotides, it is possible to obtain unprecedented sensitivity when detecting somatic mutations. We will show how the use and optimization of SuPrimers™ (DNA primers with increased specificity), BaseBlockers™ (oligos suppressing amplification of wildtype genes) and HydrolEasy probes (hydrolysis probes with increased signal-to-noise ratio and sensitivity) in combination can yield assays with a sensitivity and specificity great enough to routinely detect less than 0.05% of mutated KRAS and BRAF in a DNA samples.

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