Nucleic Acid Separation Substrate With Integrated Amplification Zone
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Solution Overview
Problem
Current methods for nucleic acid separation and amplification are complex, time-consuming, and require multiple steps, including washing processes that are not compatible with subsequent steps, especially when dealing with limited biological samples.
Innovation Solution
A method and substrate that allow for simultaneous separation and amplification of nucleic acids without a washing step by applying a sample to a substrate with a sample application zone and flowing a nucleic acid amplification reaction mixture, where the target nucleic acids are immobilized and the amplified products migrate based on molecular weight, enabling separation at a detection zone.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If multiple washing steps are performed to separate nucleic acids, then separation purity is improved, but process complexity and time consumption increase
Solution Approach 1:
The patent combines the separation and amplification steps into a single integrated process on one substrate. The substrate includes a separation zone where nucleic acids are separated by molecular weight, and an amplification zone where the separated nucleic acids are amplified. This eliminates the need for separate washing steps between separation and amplification, reducing process complexity while maintaining separation purity.
2Manufacturing precision
If multiple washing steps are performed to separate nucleic acids, then separation purity is improved, but time consumption increases
Solution Approach 1:
The patent enables continuous processing where nucleic acids are separated on the substrate and then immediately amplified in the amplification zone without interruption or washing steps in between. The reaction mixture flows continuously through the substrate, maintaining the nucleic acids in the amplification reaction mixture environment throughout the process, thus eliminating time loss associated with washing steps.
3Manufacturing precision
If washing buffer is used to remove contaminants, then purity is improved, but compatibility with subsequent amplification steps deteriorates
Solution Approach 1:
The substrate is segmented into distinct functional zones: a separation zone for separating nucleic acids by molecular weight, and an amplification zone for amplifying the separated nucleic acids. This spatial segmentation allows the separation process to occur without requiring washing steps that would be incompatible with subsequent amplification, as each zone performs its function independently on the same substrate.
4Manufacturing precision
If separate substrates are used for separation and amplification, then separation effectiveness is improved, but device complexity increases
Solution Approach 1:
The patent merges the separation substrate and amplification substrate into a single integrated substrate. The substrate contains both a separation zone for separating nucleic acids by molecular weight and an amplification zone for amplifying the separated nucleic acids. This single substrate design maintains separation effectiveness while eliminating the complexity of using multiple separate substrates and the transfer steps between them.
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 simplifies the process, reduces time, and eliminates the need for washing steps, making it suitable for small sample quantities, such as those from biopsies or forensic samples, while maintaining the integrity of the nucleic acids for downstream analysis.
Implementation Method 1
separating the target nucleic acid having a first molecular weight from the amplification product having a second molecular weight according to molecular weights of the amplified nucleic acids and target nucleic acids
Implementation Method 2
flowing a nucleic acid amplification reaction mixture across a length of the substrate through the sample application zone to amplify the target nucleic acid forming a nucleic acid amplification product
Data Source
Figure 1~2
Figure 3~4
Figure 5A~5B
AI summary
A method is provided herein, the method includes : applying a sample comprising target nucleic acids to a sample application zone of a substrate; and flowing a nucleic acid amplification reaction mixture across a length of the substrate through the sample application zone to amplify the target nucleic acid forming a nucleic acid amplification product; wherein the target nucleic acid having a first molecular weight is substantially immobilized at the sample application zone and wherein the amplification product having a second molecular weight migrates away from the sample application zone. An associated device is also provided.