Aprotic-Protic Salt DNA Stabilization at Ambient Temperature

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Solution Overview

Problem

DNA stability in aqueous media at ambient conditions is challenging due to susceptibility to hydrolysis, depurination, and denaturation, and existing preservation methods require specialized equipment or are costly, especially for long-term storage and forensic applications.

Innovation Solution

Aprotic-protic salts (APSs) are used to stabilize DNA by dissolving it in a solution comprising compounds with linked protic and aprotic centers, allowing for long-term storage at ambient temperature without the need for cryogenic preservation, enabling direct PCR amplification and reducing DNA loss during extraction and quantification steps.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If DNA is stored in aqueous media at ambient temperature, then storage cost and equipment requirements are reduced, but DNA stability and integrity deteriorate due to hydrolysis, depurination, and denaturation

Engineering Contradiction:

Improvestorage cost and equipment requirementsVSAvoidDNA stability and integrity

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

The patent introduces aprotic-protic salts as an intermediary substance between DNA and the aqueous environment. These salts form a protective complex with DNA, mediating the interaction between DNA and water to prevent hydrolysis and depurination while maintaining DNA solubility and accessibility for PCR amplification.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the chemical environment parameters by using aprotic-protic salts to modify the solvent properties. This alters the chemical reactivity of water toward DNA, reducing hydrolysis rates, and changes the physical parameters to maintain DNA stability at ambient temperature without requiring cryogenic conditions.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If DNA is stored using conventional cryogenic methods, then DNA stability is maintained, but storage cost and equipment complexity increase significantly

Engineering Contradiction:

ImproveDNA stabilityVSAvoidspecialized equipment requirements

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent replaces expensive, complex cryogenic storage infrastructure with a simple, inexpensive chemical solution (aprotic-protic salts) that can be stored in standard laboratory conditions. The chemical protector acts as a disposable or reusable additive rather than requiring ongoing energy-intensive equipment operation.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent substitutes the mechanical/physical system of cryogenic freezing with a chemical system based on aprotic-protic salt protection. Instead of using mechanical cooling equipment to maintain DNA stability, the solution uses chemical interactions to protect DNA at ambient temperature.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Ease of operation

If DNA is stored in conventional aqueous buffers, then DNA remains soluble and accessible, but DNA degradation occurs through hydrolysis and depurination

Engineering Contradiction:

ImproveDNA solubility and accessibilityVSAvoidhydrolysis, depurination, and deamination

Core Design Contradiction:

Ease of operationVSObject-affected harmful factors

Solution Approach 1:

The patent converts the harmful effect of water (which causes hydrolysis and depurination) into a beneficial effect by using aprotic-protic salts to control water's behavior. The same water molecules that would normally degrade DNA are now part of a controlled chemical environment where their reactivity is reduced, and they contribute to maintaining DNA solubility without causing degradation.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

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

The APSs maintain DNA integrity for at least two years at room temperature, reducing the need for refrigeration and enabling direct PCR amplification, thus providing a cost-effective and efficient method for preserving DNA in forensic and biotechnological applications.

Implementation Method 1

The electrostatic interaction between choline and the DNA phosphate backbone is strong

Methodology Applied

Scientific EffectElectrostatic interaction: Coulomb's Law

Implementation Method 2

dry storage of nucleic acids by exploiting the principles of anhydrobiosis

Methodology Applied

Scientific EffectAnhydrobiosis:

Implementation Method 3

hydrogen bonds between nucleotide bases, hydrogen bonds between bases and surrounding water molecules

Methodology Applied

Scientific EffectHydrogen bonding:

Implementation Method 4

base-stacking interactions between adjacent bases

Methodology Applied

Scientific EffectBase-stacking interactions: London Dispersion Force

Data Source

PatentUS11434520B2Long-term DNA preservation and storage at ambient temperature

Publication Date: 2022.09.06 FLORIDA GULF COAST UNIV BOARD OF TRUSTEES

AI summary

Aprotic-protic ionic salt (APS) compositions and methods of using aprotic-protic ionic salt compositions to stabilize nucleic acids at ambient temperatures are provided. Certain aspects provide aprotic-protic ionic salt compositions for long term storage of nucleic acids at ambient temperature in the presence of aqueous solvents.