Nucleic Acid Isolation Using Masking Solution for Chaotropic Salt Neutralization
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Solution Overview
Problem
Existing nucleic acid extraction kits struggle to efficiently isolate DNA from biological samples preserved in chaotropic salt-containing solutions, such as the DNA/RNA Shield stabilization solution, due to interference from the stabilization reagents and changes in sample constitution.
Innovation Solution
The use of a masking solution comprising C3-10 alkyl or alkenyl phenol compounds, such as eugenol, thymol, and carvacrol, in an organic solvent to neutralize the effects of chaotropic salts in the sample, allowing for effective nucleic acid isolation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a standard nucleic acid extraction kit is used on samples preserved in chaotropic salt-containing solutions, then the extraction protocol can be simple and fast, but the DNA yield is drastically reduced due to interference from stabilization reagents
Solution Approach 1:
The patent applies preliminary action by adding the masking solution containing phenolic compounds to the stabilized sample before the nucleic acid extraction process. This pre-treatment step neutralizes the chaotropic salts and masking agents in the stabilization solution, preventing them from interfering with the subsequent silica membrane-based extraction. The sample is prepared in advance by incubating with the masking solution, which restores the binding chemistry compatibility without requiring changes to the standard extraction protocol.
2Quantity of substance
If toxic phenol-containing solutions like NucleoZOL are used to neutralize chaotropic salts, then DNA yield is restored, but the method becomes highly toxic and requires special handling under a fume hood
Solution Approach 1:
The patent applies parameter changes by modifying the chemical composition of the neutralizing agent. Instead of using highly toxic phenol-containing solutions like NucleoZOL, the invention uses masking solutions containing phenolic compounds such as eugenol, thymol, or carvacrol combined with chaotropic salts. This changes the toxicity parameter while maintaining the functional ability to neutralize interference from stabilization solutions. The new reagent composition achieves the same DNA yield restoration without the severe health and safety hazards of traditional phenol-based reagents.
3Stability of the object's composition
If chaotropic salt-containing stabilization solutions are used to preserve biological samples, then sample stability is improved, but the constitution of the sample changes over time making nucleic acid extraction problematic
Solution Approach 1:
The patent applies the intermediary principle by introducing a masking solution as a mediating substance between the stabilized sample and the extraction reagents. The masking solution contains phenolic compounds that interact with and neutralize the chaotropic salts and other masking agents in the stabilization solution. This intermediary step restores compatibility between the sample matrix and the silica membrane binding chemistry, enabling efficient nucleic acid extraction without requiring changes to the standard extraction protocol.
Data Source
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AI summary
The current invention relates to the isolation of nucleic acids from sample materials present or preserved in chaotropic salt containing solutions and suitable kits therefore.