Microfluidic Device for Nucleic Acid Barcoding and Recovery
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Solution Overview
Problem
Current microfluidic devices for nucleic acid amplification and analysis are complex, fragile, and inefficient, with challenges in fluid transport and product recovery, limiting their versatility and utility in biochemical analyses.
Innovation Solution
A method and system for amplifying and tagging target nucleic acids using primers with nucleotide tags and barcode sequences, allowing for efficient amplification, barcoding, and recovery of reaction products in microfluidic devices, enabling simultaneous analysis of multiple samples and reduced operational complexity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If microfluidic devices are used for nucleic acid amplification and analysis, then conservation of reagents and samples is achieved, but device complexity and fabrication costs increase
Solution Approach 1:
The patent describes a microfluidic device that integrates multiple functions including sample lysis, nucleic acid amplification, barcoding, and product recovery in a single platform. The device can perform various biochemical analyses beyond just nucleic acid work, making it universally applicable to different assays and reducing the need for multiple specialized devices.
Solution Approach 2:
The device incorporates mechanisms for recovering reaction products after amplification and barcoding steps. This recovery capability allows for the reuse of valuable nucleic acid products and reduces waste, directly addressing the reagent conservation benefit while maintaining the integrated complex structure.
2Productivity
If microfluidic devices are miniaturized for high density analysis, then throughput increases, but device fragility and manufacturing difficulty increase
Solution Approach 1:
The microfluidic device is designed with segmented functional zones including separate chambers for lysis, amplification, and barcoding, connected by microfluidic channels. This segmentation allows for modular manufacturing approaches and simplifies the fabrication process by breaking down the complex device into manageable sections that can be manufactured and assembled more easily.
3Ease of operation
If electrical fields are used for fluid transport in microfluidic devices, then fluid control precision improves, but operational complexity increases
Solution Approach 1:
The microfluidic device incorporates passive fluid transport mechanisms such as capillary action channels and pressure-driven flow paths that eliminate the need for complex external electrical field control systems. The device structure itself provides the means for fluid transport through carefully designed channel geometries and material properties, reducing operational complexity while maintaining control precision.
4Productivity
If barcoding and tagging methods are implemented for multi-sample analysis, then analysis efficiency improves, but amplification uniformity becomes more difficult to maintain
Solution Approach 1:
The patent describes a method where barcoding and tagging are performed in a preliminary step before the main amplification reaction. By pre-tagging the nucleic acid samples with barcodes and nucleotide tags before amplification, the system enables subsequent parallel processing of multiple samples while maintaining uniform amplification conditions, thus preserving amplicon copy number uniformity across all samples.
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 method enables uniform amplification and recovery of nucleic acid products, reducing the complexity and cost of microfluidic device operations, improving the efficiency of nucleic acid analysis and sequencing processes.
Implementation Method 1
The barcode primer specifically anneals to the nucleotide tag
Implementation Method 2
The amplification mixture is subjected to amplification to produce a plurality of target amplicons
Data Source
AI summary
In certain embodiments, the present invention provides amplification methods in which nucleotide tag(s) and, optionally, a barcode nucleotide sequence are added to target nucleotide sequences. In other embodiments, the present invention provides a microfluidic device that includes a plurality of first input lines and a plurality of second input lines. The microfluidic device also includes a plurality of sets of first chambers and a plurality of sets of second chambers. Each set of first chambers is in fluid communication with one of the plurality of first input lines. Each set of second chambers is in fluid communication with one of the plurality of second input lines. The microfluidic device further includes a plurality of first pump elements in fluid communication with a first portion of the plurality of second input lines and a plurality of second pump elements in fluid communication with a second portion of the plurality of second input lines.