Nucleic Acid Isolation Using Alkylene Sulfonyl Solid Phase
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Solution Overview
Problem
Current methods for isolating nucleic acids are time-consuming, costly, and inefficient, particularly when dealing with complex biological samples, as they often require specific chaotropic salts and limited types of solid materials.
Innovation Solution
A method involving a solid phase with functional groups of structural formula (I) is used, where the sample is contacted with a first aqueous solution of pH 6-9, washed with a detergent-containing solution of similar pH, and then eluted with a low ionic strength liquid of pH 5-9 to isolate nucleic acids, allowing for a broader range of binding solutions and materials.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If existing methods are used to isolate nucleic acids, then high quality nucleic acids can be obtained, but the process is time-consuming and costly
Solution Approach 1:
The invention changes the chemical parameters of the binding solution by using kosmotropic salts (such as ammonium sulfate, sodium sulfate, magnesium sulfate) instead of traditional chaotropic salts. This parameter change enables nucleic acid binding to solid phases under different chemical conditions, simplifying the isolation process and reducing time while maintaining high nucleic acid quality through selective binding and washing
Solution Approach 2:
The invention makes the solid phase material with functional groups (carboxyl, hydroxyl, amino, sulfhydryl, or boronic acid groups) universal for binding nucleic acids under a broader range of conditions. This multi-functional approach allows the same solid phase to work with different kosmotropic salts and in the presence of detergents, reducing the need for multiple specialized reagents and steps, thereby saving time and cost
2Manufacturing precision
If existing methods are used to isolate nucleic acids, then high quality nucleic acids can be obtained, but the cost is high
Solution Approach 1:
The invention changes the cost parameter by using inexpensive kosmotropic salts (ammonium sulfate, sodium sulfate, magnesium sulfate) that are commonly available and cheaper than traditional chaotropic salts. These salts enable effective nucleic acid binding to solid phases, maintaining high quality isolation while significantly reducing reagent costs
Solution Approach 2:
The invention employs disposable solid phase materials (such as magnetic beads or solid support particles) with functional groups that can be used once and discarded. This approach eliminates the need for expensive and complex reagent systems, reducing overall isolation cost while maintaining high nucleic acid quality through effective binding and selective washing
3Manufacturing precision
If existing methods are used to isolate nucleic acids, then high quality nucleic acids can be obtained, but the procedure is complex
Solution Approach 1:
The invention simplifies the procedure by changing the binding conditions to use kosmotropic salts at physiological pH (6-9) in the presence of detergents. This parameter change eliminates the need for multiple specialized solution preparations and complex step sequences, reducing procedural complexity while maintaining high nucleic acid quality through effective binding and selective washing
Solution Approach 2:
The solid phase material with functional groups serves multiple functions: binding nucleic acids from complex biological samples containing proteins and other molecules, selective retention during washing with detergent-containing solutions, and efficient release during elution. This multi-functionality consolidates multiple steps into a simpler procedure while maintaining high isolation quality
4Manufacturing precision
If existing methods are used to isolate nucleic acids, then high quality nucleic acids can be obtained, but selectivity is limited
Solution Approach 1:
The invention expands the versatility of binding solutions by using kosmotropic salts (ammonium sulfate, sodium sulfate, magnesium sulfate) at various concentrations and combinations with detergents at physiological pH. This parameter expansion allows selective binding of nucleic acids from diverse sample types while maintaining high quality, adapting to different isolation needs without changing the fundamental methodology
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 provides a cost-effective and efficient means to obtain high-quality nucleic acids from various samples by using kosmotropic and chaotropic salts, and a wide range of solid materials, while selectively removing unwanted substances, thus improving the nucleic acid isolation process.
Implementation Method 1
contacting a sample containing nucleic acids with a solid phase in a first aqueous solution having a pH from about 6 to about 9 to provide a loaded solid phase
Implementation Method 2
washing the loaded solid phase with a second aqueous solution having a pH from about 6 to about 9 to provide a washed solid phase; wherein the second aqueous solution comprises a detergent and one or more salts
Implementation Method 3
eluting the washed solid phase with a low ionic strength liquid having a pH from about 5 to about 9 to obtain the isolated nucleic acids
Data Source
AI summary
Methods for isolating nucleic acids using a solid phase having alkylene sulfonyl-containing compounds on its surface are provided. In one embodiment, the method comprises: contacting a sample containing nucleic acids with a solid phase having alkylene sulfonyl-containing compounds on its surface in a first aqueous solution to provide a loaded solid phase; washing the loaded solid phase with a second aqueous solution to provide a washed solid phase; and eluting the washed solid phase with a low ionic strength liquid to obtain the isolated nucleic acids. Kits containing a solid phase having alkylene sulfonyl-containing compounds also are provided.