Stable Electrophoresis Medium for DNA Separation
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Solution Overview
Problem
Conventional capillary electrophoresis media containing urea suffer from chemical instability, leading to pH drift, bubble formation, and reduced shelf life, which impede the performance and stability of DNA separation during analysis.
Innovation Solution
A stable electrophoresis separation medium is developed, comprising non-crosslinked or sparsely cross-linked polymers, hydrolytically stable denaturants, and an aqueous solvent, which maintains functional stability for at least seven days at 23°C, eliminating the need for refrigeration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional electrophoresis media containing urea are used, then DNA separation is achieved, but chemical instability occurs leading to pH drift, bubble formation, and reduced shelf life
Solution Approach 1:
The patent changes the chemical parameters of the electrophoresis medium by replacing urea with alternative denaturants such as formamide, guanidinium thiocyanate, or lithium dodecyl sulfate. This parameter change eliminates the chemical instability and bubble formation associated with urea while maintaining DNA denaturation functionality, thereby extending shelf life and improving reliability
Solution Approach 2:
The patent extracts and removes urea from the electrophoresis medium formulation. By eliminating the problematic urea component entirely and substituting it with more stable denaturants, the invention resolves the chemical instability issue while preserving the essential DNA separation function
2Reliability
If refrigeration is used to maintain stability of urea-based media, then chemical degradation is reduced, but operational complexity and cost increase
Solution Approach 1:
The patent creates a disposable, room-temperature-stable electrophoresis medium that eliminates the need for refrigeration. The medium is formulated to be chemically stable under ambient conditions, allowing users to store and operate it without specialized equipment, thereby simplifying operation and reducing costs
3Reliability
If urea is used as denaturant, then DNA denaturation is achieved, but pH drift and bubble formation occur during analysis
Solution Approach 1:
The patent converts the harmful effects of urea (pH drift, bubble formation) by replacing it with denaturants that do not exhibit these problematic behaviors. The alternative denaturants maintain the beneficial DNA denaturation effect while eliminating the harmful side effects, effectively transforming the system from harmful to beneficial
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 medium provides extended shelf life and maintains DNA separation performance at room temperature, reducing the risk of degradation and ensuring consistent results over time.
Implementation Method 1
dissolved, water-soluble polymers, for instance, linear polyacrylamides, poly-N-substituted acrylamides, or poly-N,N-disubstituted acrylamides, which, at appropriate concentrations, are designed to form entangled polymer networks that physically 'sieve' DNA molecules according to their size and shape during electrophoresis
Implementation Method 2
The mediums described herein are useful for separations, such as for separations of biomolecules (including but not limited to nucleic acids, proteins, glycoproteins and glycans)
Data Source
AI summary
Provided herein is an electrophoresis separation medium comprising: (a) a non-crosslinked or sparsely cross-linked polymer or copolymer; (b) one or more denaturant compounds, in an amount sufficient to inhibit re-naturation of single stranded polynucleotides; (c) an aqueous solvent; (d) optionally, a wall-coating material suited to inhibition of electroosmotic flow; and (e) optionally, an organic water miscible solvent such as DMSO or acetonitrile, wherein the electrophoresis separation medium exhibits functional stability for at least seven days at 23° C.Also provided herein are sieving compositions, including polymer-based sieving compositions, for molecular sieving as well as related kits, devices and methods of use. Such compositions can be useful for separation of biomolecules such as nucleic acids, proteins, glycoproteins and glycans.