Abstract
Droplet digital PCR (ddPCR) has come to be regarded as the gold standard for the ultrasensitive detection and absolute quantification of closely related DNA sequences within complex mixtures. Most ddPCR assays to date, however, rely on sets of hydrolysis probes conjugated with dyes having different emission spectra to allow independent counting of rare mutant and wild-type alleles. Here, we describe a set of novel strategies that leverage the simultaneous detection and quantification of both mutant and wild-type alleles with a single hydrolysis probe. Variants of these strategies empower multiplexing and a more cost-effective approach for concurrent screening of multiple genetic variants.
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Acknowledgments
This study was supported by the Canadian Institute for Health Research (CIHR) (New Investigator award and operating grant 300738), the Terry Fox Research Institute (projects #1043 and #1021), the Natural Sciences and Engineering Research Council of Canada (Research Tools and Instruments program EQPEQ 1501), and the BC Cancer Foundation.
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Alcaide, M., Morin, R.D. (2018). Novel Multiplexing Strategies for Quantification of Rare Alleles Using ddPCR. In: Karlin-Neumann, G., Bizouarn, F. (eds) Digital PCR. Methods in Molecular Biology, vol 1768. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7778-9_16
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DOI: https://doi.org/10.1007/978-1-4939-7778-9_16
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