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Multiscale microfluidic platform with vacuum-driven chambers for automated high-volume ssDNA generation.

Anni Hu1, Yang Bu2, Yuze Liu1

  • 1Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR, China. eelyobas@ust.hk.

Lab on a Chip
|November 6, 2025
PubMed
Summary

This study introduces a novel microfluidic platform for generating large volumes of single-stranded DNA (ssDNA) using plasmonic bead-based PCR. The system efficiently processes 20 μL reactions in under 15 minutes, enabling high-throughput DNA preparation.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Molecular Biology

Background:

  • Conventional microfluidic systems face limitations in handling large fluid volumes, hindering preparative applications.
  • Efficient generation of single-stranded DNA (ssDNA) is crucial for various molecular biology techniques.
  • Plasmonic bead-based PCR offers a method for DNA amplification but requires optimization for high-volume processing.

Purpose of the Study:

  • To develop and demonstrate a multiscale microfluidic platform for automated, high-volume ssDNA generation.
  • To overcome the volume limitations of traditional microfluidic devices.
  • To achieve rapid and efficient ssDNA preparation using plasmonic PCR.

Main Methods:

  • Utilized vacuum-driven chambers (tens of microliters) integrated with microfluidic components (micromixers, microvalves).
  • Employed plasmonic thermocycling via volumetric heating with gold nanorods.
  • Incorporated magnetic beads functionalized with reverse primers for selective amplification and strand separation.

Main Results:

  • Demonstrated automated ssDNA preparation in large volumes (20 μL).
  • Achieved 20 PCR cycles in 12 minutes with high efficiency.
  • Reported 66% ssDNA recovery and 92% retention of complementary strands, indicating minimal double-stranded DNA (dsDNA) contamination.

Conclusions:

  • The developed platform enables scalable and automated high-volume ssDNA generation.
  • Vacuum-driven chambers overcome pressure-driven flow limitations in microfluidics for preparative applications.
  • This integrated system offers a robust solution for demanding molecular biology workflows requiring large ssDNA quantities.