Microfluidic Nucleic Acid Purification Device
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Solution Overview
Problem
Existing nucleic acid purification methods are cumbersome, time-consuming, and yield reproducibility depends heavily on operator skill due to manual handling and sequential application of different liquids, leading to variability in purification results.
Innovation Solution
A microfluidic device with a membrane filter and automated valves and pressure control system that allows for automated, reproducible purification of biological or chemical analytes from complex samples by controlling liquid flow and pressure differences, eliminating the need for manual manipulation and ensuring consistent contact time and flow distribution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If manual vacuum purification is used with sequential liquid application, then purification can be performed, but operator skill dependency causes yield variability and reproducibility issues
Solution Approach 1:
The system uses automated pressure control and valve actuation to perform purification steps without manual intervention. The controller automatically sequences the opening/closing of valves and application of pressure differentials to achieve consistent sample loading, washing, and elution, eliminating operator skill variability.
Solution Approach 2:
Manual vacuum control and pipetting operations are replaced by an automated control system that uses electronic pressure control and valve actuation. The controller manages the entire purification process through programmed sequences, substituting manual mechanical operations with automated electronic control.
2Reliability
If multiple sequential liquid steps are performed manually, then complete purification protocol can be executed, but process time increases and contamination risk increases
Solution Approach 1:
The system maintains continuous pressure differential control throughout the purification process, allowing sequential steps (sample loading, washing, drying, elution) to proceed without interruption. The automated valve control ensures continuous fluid flow through the filter medium, eliminating idle time between manual operations.
Solution Approach 2:
The system pre-positions valves and pressure control mechanisms in readiness for each purification step. The controller sequences valve actuation and pressure application in advance, ensuring that each liquid step is immediately ready to proceed, minimizing transition time between steps.
3Reliability
If vacuum suction is maintained for extended periods after liquid flow, then complete liquid removal is achieved, but process time increases
Solution Approach 1:
The system uses pressure sensors to monitor the pressure differential across the filter medium in real-time. When the pressure differential indicates that liquid flow has ceased and the filter is sufficiently dry, the controller automatically adjusts or terminates the vacuum suction, eliminating the need for fixed extended vacuum maintenance times.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The automated microfluidic device achieves reproducible nucleic acid purification independent of operator skill, reducing contamination and increasing yield consistency compared to manual methods, while allowing seamless integration with downstream applications like qPCR.
Implementation Method 1
nucleic acid attraction for silica surfaces is promoted by a high concentration of chaotropic salts
Implementation Method 2
The Boom method uses a chaotropic salt solution to denaturalize the biological sample and make it go through the filter using centrifugal forces to promote DNA and RNA adsorption onto the silica surface
Implementation Method 3
make it go through the filter using centrifugal forces to promote DNA and RNA adsorption onto the silica surface
Implementation Method 4
one or more washes with ethanolic buffers are performed to get rid of the chaotropic salts and other biological impurities while keeping nucleic acids bound
Implementation Method 5
after getting rid of ethanol (with a high speed spin), nucleic acids need to get rehydrated
Implementation Method 6
nucleic acids need to get rehydrated using an elution buffer (water or low salt buffer). Rehydration promotes unbinding of the DNA and RNA from the silica surface
Data Source
Figure 1~2A
Figure 2B~2C
Figure 2D~2E
AI summary
A microfluidic device is disclosed having an enclosed chamber containing a filter for purifying biological or chemical analytes from a complex biological sample, said chamber housing a plurality of ports in addition to said filter, as follows: a first port enabling gas communication of the chamber with a vacuum generator, via a first flow path; a second port enabling liquid communication of the chamber with one or more reservoirs, via a second flow path; a third port enabling gas and liquid communication of the chamber with both one or more receiving containers and a vacuum generator, via a third flow path; and a filter located between the third port and both the first and second port, so that a fluid entering the chamber through the first and/or second port and exiting the chamber through the third port flows through the filter. The invention also relates to a method using the microfluidic device.