Microfluidic Nucleic Acid Desorption via Segmented Elution
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Solution Overview
Problem
Current methods for desorbing nucleic acids from samples often result in high concentrations of inhibitory components, which can interfere with subsequent analysis, and existing technologies do not efficiently reduce these inhibitors while maximizing nucleic acid yield.
Innovation Solution
A method utilizing a microfluidic system where a solid phase is rinsed multiple times with an elution buffer to elute nucleic acids, employing a micropump to control precise elution volumes and flow directions, reducing inhibitor concentrations and optimizing nucleic acid recovery through fractionation and mixing of eluate fractions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If a single elution step is used to maximize nucleic acid recovery, then nucleic acid yield is improved, but inhibitor concentration remains high and interferes with subsequent analysis
Solution Approach 1:
The elution process is divided into multiple sequential steps with different elution buffers. The first elution step uses a buffer optimized for nucleic acid recovery, while subsequent steps use buffers optimized for inhibitor removal. This segmentation allows independent optimization of each step's function, resolving the contradiction between maximizing yield and minimizing inhibitors.
Solution Approach 2:
The patent implements continuous elution steps without interrupting the process. Each elution step immediately follows the previous one, with the eluate from one step becoming the input for the next. This continuous action ensures that nucleic acids are progressively purified while maintaining high recovery, preventing the contradiction from manifesting.
2Object-affected harmful factors
If multiple elution steps are performed to reduce inhibitor concentration, then inhibitor concentration is reduced, but processing time and complexity increase
Solution Approach 1:
Each elution step uses buffers with different chemical parameters (pH, salt concentration, organic solvent content) specifically optimized for that step's purpose. The first buffer prioritizes nucleic acid solubility, while subsequent buffers prioritize inhibitor removal. This parameter optimization allows each step to be highly efficient, reducing the total time required despite multiple steps.
Solution Approach 2:
The patent extracts inhibitors from the eluate in sequential steps, with each buffer specifically designed to remove particular types of inhibitors. By targeting and removing inhibitors in dedicated steps rather than attempting simultaneous removal, the process achieves thorough purification more efficiently than a single complex step would allow.
3Productivity
If large elution volumes are used to ensure complete nucleic acid elution, then elution efficiency is improved, but inhibitor concentration increases and nucleic acid concentration decreases
Solution Approach 1:
The total elution volume is segmented across multiple steps rather than delivered in a single large volume. Each step uses a smaller, optimized volume that achieves sufficient elution efficiency while concentrating the nucleic acid and removing inhibitors progressively. This segmentation resolves the contradiction between complete elution and inhibitor concentration.
Solution Approach 2:
The elution process uses periodic application of different buffers in a cycle: first buffer for nucleic acid release, second buffer for inhibitor removal, third buffer for final purification. This periodic action with varying buffer compositions achieves complete elution efficiency while systematically reducing inhibitor concentration at each cycle.
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
This approach effectively reduces inhibitory substance concentrations, allowing for successful enzymatic detection and amplification of nucleic acids, even at low concentrations, by achieving maximum elution efficiency and precise control over elution processes in the microfluidic system.
Implementation Method 1
the lysate produced by lysis is often mixed with a binding buffer and brought into contact with a solid matrix, such as a silica filter, whereby the nucleic acids adsorb to the filter
Implementation Method 2
a solid phase in a microfluidic system is repeatedly rinsed with an elution buffer in order to elute nucleic acids bound to the solid phase in the microfluidic system from the solid phase
Data Source
Figure 1~2
Figure 3
AI summary
The invention relates to a method for the desorption of nucleic acids from a sample. In order to simplify the desorption of nucleic acids from the sample, a solid phase (16) is repeatedly rinsed with an elution buffer in a microfluidic system (1), in order to elute nucleic acids bonded to the solid phase (16) from the solid phase (16) in the microfluidic system (1).