Absolute quantification of nucleic acid on digital microfluidics platform based on superhydrophobic-superhydrophilic micropatterning
SENSORS AND ACTUATORS B-CHEMICAL(2024)
摘要
Digital microfluidics (DMF) shows great potential in the manipulation of individual droplets. However, the limited number of electrode control wirings for droplet control intrinsically restrains the massive generation of pL/nL individual droplets, fundamentally hindering the realization of high throughput applications on DMF such as digital nucleic acid amplification and analysis. This work demonstrates on-chip high throughput digital loopmediated isothermal amplification (dLAMP) on a DMF platform based on a functional substrate with superwettability contrast. Exploiting the passive dispensing technique as empowered by electrowetting on dielectric (EWOD) effect, a total of 1818 individual droplets with high uniformity are generated on the super-hydrophilic microarray within 1 min. Separation within the oil phase without tubes and valves can avoid evaporation, ensuring a precise total reaction volume. Absolute quantification of lambda DNA with a dynamic range of 1-1000 copies/mu L is achieved by counting the amount of positive microdroplets after on-chip incubation, demonstrating the strong potential for rapid, low-cost, reliable, and quantitative nucleic acid analysis with high accuracy.
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关键词
High-throughput,Digital Microfluidics,DLAMP,Super-wettability
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