Journal
ANALYTICAL CHEMISTRY
Volume 90, Issue 8, Pages 5329-5334Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b00463
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Funding
- Tokyo Metropolitan University
- National Key R&D Program of China [2017YFC0906800]
- National Natural Science Foundation of China [21435002, 21621003]
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We report on the development of a novel and flexible online digital polymerase chain reaction (dPCR) system. The system was composed of three parts: an inkjet for generating the droplets, a coiled fused-silica capillary for thermal cycling, and a laser-induced fluorescence detector (LIFD) for positive droplet counting. Upon inkjet printing, monodisperse droplets were continuously generated in the oil phase and then introduced into the capillary in the form of a stable dispersion. The droplets containing one or zero molecules of target DNA passed through the helical capillary that was attached to a cylindrical thermal cycler for PCR amplification, resulting in the generation of fluorescence for the DNA-positive droplet. After 36 PCR cycles, the fluorescence signal intensity was detected by laser-induced fluorescence located at the downstream of the capillary, followed by a positive/negative counting. The present system was successfully applied to the absolute quantification of the HPV sequence in Caski cells with dynamic ranges spanning 4 orders of magnitude.
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