Microdroplet PCR in Microfluidic Chip Based on Constant Pressure Regulation

Area of Science:

• Biotechnology
• Analytical Chemistry
• Microfluidics

Background:

• Microdroplet Polymerase Chain Reaction (PCR) is crucial for high-throughput genetic analysis.
• Controlling microdroplet behavior, such as movement, fragmentation, and bubble formation, is essential for reliable PCR.
• Existing microfluidic PCR methods face challenges with bubble generation and sample integrity.

Purpose of the Study:

• To develop a device and method for constant pressure regulation in microdroplet PCR.
• To optimize microdroplet generation, movement, and prevent bubble formation in microfluidic chips.
• To enable sensitive and quantitative detection of human genes using microdroplet PCR.

Main Methods:

• Utilized an air source device for precise pressure regulation within the microfluidic chip.
• Generated approximately 50,000 water-in-oil microdroplets (87 μm diameter) from a 20 μL sample in 3 minutes.
• Integrated microdroplet generation and PCR amplification, ensuring a bubble-free environment.

Main Results:

• Achieved consistent microdroplet generation and close arrangement within the chip without air bubbles.
• Demonstrated quantitative detection of human genes with a wide linear range (10^1 to 10^5 copies/μL, R² = 0.999).
• Highlighted advantages including high pollution resistance, avoidance of microdroplet fragmentation/integration, reduced human interference, and standardized results.

Conclusions:

• The developed constant pressure regulation system effectively controls microdroplet PCR in microfluidic chips.
• This technology facilitates bubble-free microdroplet generation and PCR amplification.
• Microdroplet PCR devices with constant pressure regulation show significant promise for accurate nucleic acid quantification.