Nucleic Acid Adsorption Using Chaotropic Agent Concentration
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Solution Overview
Problem
Current methods for isolating nucleic acids from biological samples are inefficient due to the presence of degrading enzymes and contaminants, and often require the use of alcohol, which is hazardous, and are not suitable for processing large quantities.
Innovation Solution
A two-step lysis and binding method using an aqueous lysis buffer with a chaotropic agent, where the concentration of the chaotropic agent is increased to enhance nucleic acid adsorption to a solid phase, such as silica or glass, allowing for effective separation and elution of nucleic acids without the need for alcohol.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If alcohol is used for nucleic acid precipitation, then nucleic acid isolation efficiency is improved, but safety hazards increase due to flammability
Solution Approach 1:
The invention changes the chemical parameter of the precipitation agent from flammable alcohol to non-flammable salts (such as sodium chloride, ammonium acetate, or ammonium sulfate). This substitution maintains the precipitation function while eliminating the safety hazard associated with alcohol flammability.
2Productivity
If degrading enzymes are present during lysis, then nucleic acid degradation occurs, but complete cell lysis is required for efficient nucleic acid release
Solution Approach 1:
The invention applies preliminary anti-action by adding protease inhibitors to the lysis buffer before cell lysis occurs. This prevents degrading enzymes from attacking the nucleic acids during the lysis process, thereby protecting nucleic acid integrity while still allowing complete cell lysis for efficient nucleic acid release.
Solution Approach 2:
The invention uses protease inhibitors as intermediaries that mediate between the degrading enzymes released during lysis and the nucleic acids. These inhibitors bind to the degrading enzymes, preventing them from degrading the nucleic acids, thus protecting the target molecules during the necessary cell lysis process.
3Productivity
If chaotropic agent concentration is increased to enhance nucleic acid adsorption, then binding efficiency is improved, but solubility of other components may be affected
Solution Approach 1:
The invention optimizes the concentration parameter of chaotropic agents in the binding buffer to achieve maximum nucleic acid adsorption efficiency. By carefully controlling the chaotropic agent concentration, the invention enhances nucleic acid binding to the solid phase while minimizing the precipitation of other soluble components, thus maintaining a balance between binding efficiency and component solubility.
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 more efficient and safer process for isolating nucleic acids, reducing the risk of enzyme degradation and enabling the processing of larger sample quantities while avoiding the use of flammable substances like alcohol.
Implementation Method 1
mixing the lysis buffer with the biological sample, whereby the concentration of the chaotropic agent in the mixture is between 1 M and 4 M... contacting the mixture of step (c) with the solid phase, thereby adsorbing the nucleic acid from the mixture to the solid phase
Data Source
AI summary
The present invention is directed to a method for adsorbing, i.e. non-covalently binding, nucleic acids to a solid phase using a two-step procedure. Furthermore, the present invention pertains to a method for isolating nucleic acids from a biological sample. In the first step of the procedure, lysis is effected by mixing the biological sample with an aqueous lysis buffer containing a chaotropic agent and incubating the mixture; in the second step, the concentration of the chaotropic agent in the mixture is increased and the mixture is contacted with the solid phase, whereby the nucleic acids in the liquid phase is adsorbed to the solid phase.