One-Tube Nucleic Acid Isolation Buffer for Rapid PCR
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Solution Overview
Problem
Conventional nucleic acid isolation methods using columns are complex, time-consuming, and require multiple steps, specialized equipment, and significant manpower, making them unsuitable for rapid onsite diagnosis, especially in emergencies like viral outbreaks.
Innovation Solution
A one-step nucleic acid isolation buffer composition comprising ethanol, SDS, sodium chloride, EDTA, and Tris-HCl, which allows for direct immobilization of nucleic acids on a membrane filter, enabling column-based one-tube PCR without the need for centrifugation, washing, or drying processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional column-based nucleic acid isolation method is used, then nucleic acid purification is achieved, but the process becomes complex and time-consuming requiring multiple steps and specialized equipment
Solution Approach 1:
The patent combines multiple isolation steps (lysis, binding, washing, elution) into a single buffer solution called one-tube buffer. This buffer contains all necessary components (detergent, salt, chaotropic agent, buffer) to perform the complete isolation process in one tube, eliminating the need for multiple separate buffers and steps while maintaining purification quality
Solution Approach 2:
The one-tube buffer serves multiple functions simultaneously: it acts as a lysis buffer to break down cells, a binding buffer to facilitate nucleic acid attachment to the column, and contains all necessary washing and elution components. This multi-functional design simplifies the overall process while maintaining effectiveness
2Reliability
If conventional column-based nucleic acid isolation method is used, then nucleic acid is isolated, but significant time and manpower are required for multiple processing steps
Solution Approach 1:
The one-tube buffer is pre-formulated with all necessary components for the complete isolation process. Users only need to add the sample to the pre-prepared buffer and column assembly, eliminating the need to prepare multiple separate buffers and perform multiple manual transfer steps, thus saving significant time while maintaining isolation effectiveness
Solution Approach 2:
The one-tube buffer system is designed to perform all isolation functions automatically within a single tube. The buffer components work synergistically to lyse cells, bind nucleic acids, and resist contamination without requiring external intervention for each step, reducing both time and manpower requirements
3Reliability
If conventional column-based nucleic acid isolation method is used, then nucleic acid purification is achieved, but multiple washing processes increase the required time
Solution Approach 1:
The patent merges the washing function into the one-tube buffer by incorporating washing agents and contaminants-binding substances directly into the buffer formulation. This eliminates the need for separate washing steps while maintaining nucleic acid purity through the integrated buffer composition
4Reliability
If conventional column-based nucleic acid isolation method is used, then nucleic acid is obtained, but specialized equipment and tools are required making onsite use difficult
Solution Approach 1:
The patent extracts the need for specialized equipment by designing a system that works with simple, universally available materials. The one-tube buffer and column assembly can be used with basic laboratory or even field equipment, removing the dependency on complex specialized instruments while maintaining extraction quality
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 simplifies the nucleic acid isolation process, reducing the risk of contamination and loss, eliminating the need for specialized equipment, and enabling rapid, cost-effective onsite PCR for diagnostic purposes.
Implementation Method 1
A one-step nucleic acid isolation buffer composition comprising ethanol, SDS, sodium chloride, EDTA, and Tris-HCl, which allows for direct immobilization of nucleic acids on a membrane filter
Implementation Method 2
A one-step nucleic acid isolation buffer composition comprising ethanol, SDS, sodium chloride, EDTA, and Tris-HCl
Data Source
AI summary
The present invention relates to a nucleic acid isolation buffer composition, a nucleic acid isolation kit in which a membrane filter comprising same is provided, and a nucleic acid isolation method, wherein the nucleic acid isolation buffer composition has excellent compatibility with membrane filters of various materials, and enables the excellent isolation of nucleic acids from various samples, and, particularly, a nucleic acid isolation kit comprising same enables column-based one-tube PCR, and a nucleic acid isolation method using same enables the direct application of the membrane filter, on which nucleic acids are immobilized, to PCR without a separate elution process. Therefore, the nucleic acid isolation composition, the kit and the isolation method according to the present invention can be effectively used in rapid diagnosis onsite.