High-throughput analysis of DNA fragments using a miniaturized CE system combined with a slotted-vial array sample...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Molecular Biology

Background:

• High-throughput analysis of DNA fragments is crucial for various applications.
• Existing microfluidic capillary electrophoresis (CE) systems face challenges in automated sample introduction and low sample consumption.

Purpose of the Study:

• To develop and evaluate an automated nanoliter sample introduction system integrated with a liquid-core waveguide (LCW)-based microfluidic CE system.
• To achieve high-throughput and sensitive analysis of DNA fragments with minimal sample volume.

Main Methods:

• An automated nanoliter sample introduction system utilizing a motor-driven plate with slotted vials was developed.
• A 7 cm LCW capillary served as both the sample probe and separation channel.
• Laser-induced fluorescence (LIF) detection with a diode laser and photomultiplier tube (PMT) was employed.

Main Results:

• Baseline separation of 11 DNA fragments from PhiX174-HaeIII digest was achieved.
• A high throughput of 33 samples per hour was demonstrated.
• Exceptional theoretical plate number (7.3x10^6/m) and low detection limit (0.4 ng/microL for 603 bp fragment) were obtained.
• Automated sample introduction consumed only 0.3 nL per cycle with 2.2% RSD for peak height.

Conclusions:

• The integrated system enables highly efficient, automated, and sensitive analysis of DNA fragments.
• This approach significantly reduces sample consumption and increases throughput for CE-based DNA analysis.
• The system holds promise for applications requiring rapid and precise DNA fragment quantification.