RNA Stabilization Composition Using Anionic Detergent and Buffer

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Solution Overview

Problem

Current methods for collecting, preserving, and analyzing RNA from bodily fluids are hindered by RNA's labile nature, requiring immediate freezing or refrigeration, which is impractical for remote or large-scale sample collection, and existing stabilization methods like cationic compounds are unsatisfactory for long-term storage at room temperature.

Innovation Solution

A composition comprising an anionic detergent and a buffering agent at a pH of 5 to 8.2, which stabilizes RNA at room temperature, combined with a heating step above 50°C to inhibit ribonuclease activity and facilitate extraction.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If RNA samples are stored at room temperature using conventional methods, then storage convenience is improved, but RNA stability deteriorates due to degradation by ribonucleases and chemical hydrolysis

Engineering Contradiction:

Improvestorage convenienceVSAvoidRNA stability

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The invention changes the chemical parameters of the storage environment by using an anionic detergent (like SDS) to denature ribonucleases and a buffering agent to maintain pH between 5-8.2, creating conditions where RNA is stable at room temperature without requiring freezing or refrigeration

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The anionic detergent acts as an intermediary substance that binds to and inactivates ribonucleases, preventing them from degrading RNA. The buffering agent serves as another intermediary that maintains optimal pH conditions for RNA stability

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If RNA samples are frozen or refrigerated for storage, then RNA stability is improved, but storage cost and complexity increase due to equipment requirements

Engineering Contradiction:

ImproveRNA stabilityVSAvoidstorage equipment requirements

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The invention replaces expensive, complex refrigeration and freezing equipment with a simple, disposable chemical stabilization solution that can be stored at room temperature, eliminating the need for freezers and refrigerators in remote or field settings

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

By changing the chemical environment through anionic detergent and buffering agents, the invention shifts RNA stability from being temperature-dependent to being chemically protected, allowing room temperature storage without specialized equipment

Inventive Principle:

Principle #35Parameter changes

3Reliability

If cationic compounds are used for RNA stabilization, then ribonuclease inhibition is improved, but RNA extraction efficiency deteriorates due to interference with downstream applications

Engineering Contradiction:

Improveribonuclease inhibitionVSAvoidRNA extraction efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

Instead of using cationic compounds (positive charge) for ribonuclease inhibition, the invention inverts the approach by using anionic detergents (negative charge) like SDS, which effectively denature ribonucleases while being removable and not interfering with subsequent RNA extraction and downstream applications

Inventive Principle:

Principle #13The other way round (Inversion)

4Reliability

If rapid transportation of RNA samples is implemented, then sample degradation is reduced, but cost and feasibility deteriorate for remote locations and large-scale collection

Engineering Contradiction:

Improvesample integrityVSAvoidcollection feasibility

Core Design Contradiction:

ReliabilityVSEase of operation

Solution Approach 1:

The invention applies preliminary stabilization action by adding the anionic detergent and buffering agent composition to RNA samples at the point of collection, protecting them from degradation before transportation begins, thereby eliminating the need for rapid transport to maintain sample integrity

Inventive Principle:

Principle #10Preliminary action

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 allows for prolonged storage and stable extraction of RNA from bodily fluids at room temperature, maintaining RNA integrity for extended periods without the need for refrigeration, enabling convenient and cost-effective RNA analysis.

Implementation Method 1

anionic detergent...inhibit ribonuclease activity

Methodology Applied

Scientific EffectDenaturation:

Implementation Method 2

a buffering agent at a pH of 5 to 8.2, which stabilizes RNA at room temperature

Methodology Applied

Scientific EffectpH buffering:

Implementation Method 3

a heating step above 50°C to inhibit ribonuclease activity and facilitate extraction

Methodology Applied

Scientific EffectThermal inhibition of enzyme activity:

Data Source

PatentUS10000795B2Stabilizing compositions and methods for extraction of ribonucleic acid

Publication Date: 2018.06.19 DNA GENOTEK

AI summary

The present invention provides a composition and method for stabilizing ribonucleic acid (RNA) from biological samples such that the ribonucleic acid within the sample remains stable at room temperature. The composition comprises an anionic detergent and a buffering agent at a pH of about 5 to about 8.2 and is used in methods for extracting and storing ribonucleic acid from the biological sample.