Nucleic Acid Extraction Kit Using Potassium Salt SDS Precipitation
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Solution Overview
Problem
Current nucleic acid extraction methods are time-consuming, require expensive and toxic chemicals, and need sophisticated equipment and laboratory settings, making them unsuitable for rapid, inexpensive, and universal extraction from complex samples, especially in non-laboratory settings such as during pandemics or field testing.
Innovation Solution
A kit and method using a lysis buffer with SDS concentrations between 1% to 25%, potassium salts between 0.1 M to 5.0 M, and zinc or copper salts between 0.5 M to 5.0 M, combined with a filter of 1 μm to 10 μm pore diameter, allowing for direct extraction of nucleic acids without additional purification steps, compatible with various sample types.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If SDS-based extraction methods are used, then nucleic acids can be released from cells, but the presence of SDS inhibits subsequent PCR amplification
Solution Approach 1:
The invention extracts and removes SDS from the reaction mixture by adding potassium salts, which cause SDS to precipitate as SDS-potassium complexes. This separation allows the nucleic acids to remain in solution while the detergent is removed, resolving the contradiction between effective lysis and PCR compatibility
Solution Approach 2:
Potassium salts serve as an intermediary substance that mediates between the lysis step (requiring SDS) and the amplification step (requiring SDS-free conditions). The potassium ions facilitate SDS precipitation without directly interacting with or inhibiting the nucleic acids or subsequent PCR reactions
2Manufacturing precision
If traditional purification steps are included, then nucleic acid purity is improved, but the extraction process becomes time-consuming and requires sophisticated equipment
Solution Approach 1:
The invention merges the lysis and purification steps into a single integrated process. The addition of potassium salts simultaneously achieves SDS removal and nucleic acid concentration in one step, eliminating the need for separate purification steps and reducing overall processing time
Solution Approach 2:
The method uses simple, inexpensive reagents (potassium salts) that can be easily disposed of after a single use, replacing expensive, complex purification equipment. This approach prioritizes simplicity and ease of disposal over reusability, suitable for point-of-care settings
3Quantity of substance
If conventional extraction methods are used, then nucleic acids can be extracted, but toxic chemicals and sophisticated equipment are required
Solution Approach 1:
The invention replaces toxic, expensive chemicals and equipment with simple, inexpensive potassium salts that require no special handling or disposal procedures. The method uses readily available materials that pose minimal harm to operators and the environment
Solution Approach 2:
The extraction system becomes self-sufficient by using inherently safe reagents that do not require sophisticated equipment for handling, storage, or disposal. The potassium salt-based system performs purification functions without needing external support systems or specialized infrastructure
4Productivity
If rapid extraction is implemented without purification steps, then processing time is reduced, but extraction accuracy and purity may be compromised
Solution Approach 1:
The invention combines lysis and purification into a single step by adding potassium salts that simultaneously precipitate SDS and concentrate nucleic acids. This merged process achieves both speed and accuracy that would otherwise require separate sequential steps
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
Enables rapid and inexpensive extraction of nucleic acids from complex samples, eliminating the need for toxic chemicals and laboratory equipment, suitable for diverse sample types, and compatible with subsequent amplification steps, facilitating point-of-care testing and universal application.
Implementation Method 1
A frequently used chemical lysis method is based on the use of sodium dodecyl sulfate (or SDS), a strong ionic detergent and surfactant, which is usually combined with proteases for even more efficient lysis. This chemical method based on the use of SDS allows membranes or walls to burst and to release nucleic acids.
Implementation Method 2
a filter having a pore diameter ranging from about 1 μm to about 10 μm
Data Source
AI summary
A kit and method for extracting nucleic acids from complex samples. The kit includes: (i) a lysis buffer having a concentration of SDS ranging from about 1% to about 25%; (ii) a buffer having a concentration of a potassium salt of ranging from about 0.1 M to about 5.0 M; (iii) a buffer having a concentration of a zinc and/or copper salt of ranging from about 0.5 M to about 5.0 M; (iv) a filter having a pore diameter ranging from about 1 μm to about 10 μm; and optionally, a member selected from one or more syringe(s), one or more reaction tube(s), an instruction guide, and any combination thereof.