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. 2021 Jul 12;93(29):9974–9979. doi: 10.1021/acs.analchem.1c00877

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© 2021 The Authors. Published by American Chemical Society

Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).

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Vortex emulsification qPCR readout enables accurate vddPCR. (a) The DNA sample is added to a tube, oil with stabilizing surfactant is introduced, and the mixture emulsified by vortexing. (b) Vortexed emulsions are thermally cycled. An aliquot is amplified with TaqMan probes to enable visualization. (c) Size distribution of the vortex emulsified droplets obtained by imaging (n = 1323). (d) qPCR readout of vddPCR allows accurate quantitation of targets over a range of concentrations. Microfluidic ddPCR, due to the uniformity of the droplets, shows slightly better accuracy and consistency for targets below 20 molecules compared to vddPCR. The droplet reader quantitates microfluidic ddPCR but requires a specialized instrument.