Size Selective DNA Isolation Using Unmodified Silicon
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Solution Overview
Problem
Current methods for size selective isolation of DNA molecules, particularly for next-generation sequencing, are time-consuming and require costly surface modifications of solid phases, limiting their efficiency and scalability.
Innovation Solution
A poly(alkylene oxide) polymer-based method using polyethylene glycol, an alkali metal salt, and a divalent cation in a binding mixture with an unmodified silicon-containing surface, allowing for efficient precipitation and recovery of DNA molecules above a certain size cut-off, independent of surface modifications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If surface modifications are applied to solid phases for size selective DNA isolation, then DNA binding efficiency is improved, but manufacturing complexity and cost increase
Solution Approach 1:
The invention extracts and removes the surface modification step from the solid phase preparation process. By using an unmodified silicon-containing surface, the complex and costly surface modification procedure is eliminated entirely, while DNA binding efficiency is maintained through the specific chemical composition of the binding mixture containing polyethylene glycol, alkali metal salt, and divalent cation.
Solution Approach 2:
The invention replaces expensive, modified solid phases with a simple, unmodified silicon-containing surface that can be used as a disposable or single-use component. The cost and complexity of modifying solid phases is eliminated by using a basic silicon surface in combination with a specifically formulated binding mixture.
2Manufacturing precision
If conventional size selective isolation methods are used, then DNA separation is achieved, but processing time increases
Solution Approach 1:
The invention changes the chemical parameters of the binding mixture by incorporating polyethylene poly(alkylene oxide) polymer, alkali metal salt, and divalent cation in specific concentrations. This parameter optimization enables faster DNA binding kinetics while maintaining size separation precision, thereby reducing overall processing time without compromising separation quality.
3Reliability
If polyethylene glycol and optimized binding conditions are used with unmodified silicon surface, then DNA recovery rate is improved, but binding mixture complexity increases
Solution Approach 1:
The invention creates a composite binding system by combining polyethylene glycol, alkali metal salt, and divalent cation in a binding mixture. This composite formulation works synergistically with the unmodified silicon-containing surface to enhance DNA recovery rates while the components remain chemically simple and easily obtainable.
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 method enhances DNA recovery rates and simplifies the process by using a standard silicon surface, improving the yield and cost-effectiveness of size selective DNA isolation, particularly for removing adapter monomers and dimers in sequencing libraries.
Implementation Method 1
a poly(alkylene oxide) polymer based method using polyethylene glycol, an alkali metal salt, and a divalent cation in a binding mixture with an unmodified silicon-containing surface, allowing for efficient precipitation and recovery of DNA molecules above a certain size cut-off
Implementation Method 2
binding precipitated DNA molecules to a solid phase having an unmodified silicon containing surface
Data Source
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AI summary
The present invention provides a poly(alkylene oxide) polymer based size selective DNA isolation method for isolating DNA molecules having a size above a certain cut-off value from a DNA containing sample, comprising (a) preparing a binding mixture comprising the DNA containing sample, at least one poly(alkylene oxide) polymer and at least one divalent cation, wherein said binding mixture has a p H that lies in the range of 8 to 10 and binding precipitated DNA molecules to a solid phase having an unmodified silicon containing surface, thereby providing a solid phase having bound thereto DNA molecules having a size above the cut-off value, wherein under the used binding conditions DNA molecules having a size which is less than the cut-off value substantially do not bind to the solid phase; (b) separating the bound DNA molecules from the remaining sample; - optionally washing the bound DNA molecules; and - optionally eluting the bound DNA molecules from the solid phase. Said method allows the size selective purification of target DNA molecules and is particularly suitable for sequencing applications. Moreover, a kit is provided.