Tunable nanofluidic device for digital nucleic acid analysis

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Diagnostics

Background:

• Nano/microfluidic devices offer rapid nucleic acid (NA) detection but face challenges in sample loading and NA retention for enzymatic reactions.
• Current methods often require pre-treatment or external actuators, limiting their efficiency and applicability.

Purpose of the Study:

• To develop a nanofluidic device with real-time confinement control for efficient NA capture and enzymatic processing.
• To demonstrate the loading, retention, and enzymatic exposure of target NAs within digital nanocavities.

Main Methods:

• Utilized a nanofluidic device with pneumatic actuation for real-time control of membrane-based confinement.
• Demonstrated target NA loading into digital nanocavities and subsequent enzymatic access.
• Employed loop-mediated isothermal amplification and rolling circle amplification for DNA amplification within nanocavities.

Main Results:

• Successfully loaded target nucleic acids into digital nanocavities.
• Achieved retention and simultaneous exposure of captured NAs to enzymes and co-factors.
• Demonstrated isothermal amplification of single-stranded DNA (M13mp18 plasmid vector) within the nanocavities.

Conclusions:

• The developed nanofluidic device effectively addresses NA loading and retention challenges in confined volumes.
• This technology shows promise for advanced, pre-treatment-free nucleic acid diagnostic applications.