Flow Cell DNA Capture Using Thermoreversible Methyl Cellulose
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Solution Overview
Problem
Existing methods for nucleic acid analysis, particularly DNA capture and concentration, suffer from inefficiencies leading to significant sample loss and inadequate purification, especially when dealing with diverse DNA sizes and types.
Innovation Solution
A method utilizing methyl cellulose as a thermally reversible precipitating polymer to form and release DNA complexes under controlled temperature conditions, enabling capture, concentration, and purification of DNA by forming and detangling DNA-methyl cellulose complexes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of substance
If traditional DNA capture methods are used, then DNA can be captured for analysis, but significant sample loss occurs and purification is inadequate
Solution Approach 1:
The patent utilizes temperature as a controllable parameter to change the solubility characteristics of methyl cellulose. By heating the sample fluid to above the gelation temperature, methyl cellulose precipitates and forms complexes with DNA, enabling capture. By cooling below the gelation temperature, the complexes dissolve and release DNA, enabling purification. This parameter-based control resolves the contradiction by providing both efficient capture and effective release without significant sample loss.
Solution Approach 2:
The patent exploits the phase transition of methyl cellulose between soluble and insoluble states at different temperatures. Above the gelation temperature, methyl cellulose transitions to an insoluble state that complexes with DNA for capture. Below the gelation temperature, it transitions back to a soluble state that releases DNA for purification. This phase transition mechanism enables both high capture efficiency and effective purification, resolving the technical contradiction.
2Quantity of substance
If DNA concentration is increased for analysis, then analysis sensitivity improves, but sample loss increases with traditional methods
Solution Approach 1:
The methyl cellulose-DNA complex system is self-service in that the same reagent system used for capture automatically enables release and purification through temperature cycling. The complex formation at high temperature concentrates DNA, while cooling automatically releases the DNA in purified form, eliminating the need for separate recovery steps that cause sample loss. This self-service mechanism achieves both concentration and minimal loss.
3Adaptability or versatility
If diverse DNA sizes are processed, then comprehensive analysis coverage is achieved, but separation and purification become more difficult
Solution Approach 1:
The patent segments the processing of diverse DNA molecules by size through the physical properties of methyl cellulose-DNA complexes. Larger DNA molecules form more extensive complexes with methyl cellulose, while smaller molecules form fewer complexes. This natural segmentation during complex formation, combined with temperature-controlled dissolution, enables size-based separation without complex equipment, resolving the contradiction between comprehensive coverage and process simplicity.
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
Maximizes DNA loading for analysis by minimizing sample loss and effectively separating different DNA sizes, enhancing purification efficiency and seeding on a flow cell surface.
Implementation Method 1
heating the sample fluid to at least a gelation temperature of the methyl cellulose, thereby forming DNA-methyl cellulose complexes in the aqueous carrier
Implementation Method 2
heating the sample fluid to at least a gelation temperature of the methyl cellulose
Implementation Method 3
a filter positioned in the temperature controlled flow channel, the filter i) to block concentrated deoxyribonucleic acid (DNA)-methyl cellulose complexes generated in the temperature controlled flow channel
Implementation Method 4
cooling the sample fluid to below the gelation temperature of the methyl cellulose, thereby detangling the concentrated DNA-methyl cellulose complexes to release the DNA sample and the methyl cellulose
Data Source
AI summary
An example of a kit includes a flow cell assembly. The flow cell assembly includes a reaction chamber, a temperature controlled flow channel in selective fluid communication with an inlet of the reaction chamber, and a filter positioned in the temperature controlled flow channel. The reaction chamber includes depressions separated by interstitial regions and capture primers attached within each of the depressions. The filter is i) to block concentrated biological sample-polymer complexes generated in the temperature controlled flow channel at a first temperature, and ii) to allow passage of concentrated biological sample and polymer released from the complexes in the temperature controlled flow channel at a second temperature.