RNA Stabilization Buffer Composition for Room-Temperature Storage
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Biological samples, particularly RNA within them, degrade over time, especially when stored at room temperature or subjected to thermal excursions, necessitating specialized equipment and processes that can damage the samples.
Innovation Solution
A stabilization buffer comprising guanidine hydrochloride, TRITON X-100, EDTA, citric acid, and sodium citrate at specific concentrations, maintaining a pH less than 4.5, effectively stabilizes biological samples, including RNA, at room temperature for extended periods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If biological samples are stored at room temperature for extended periods, then storage convenience is improved, but RNA degradation increases
Solution Approach 1:
The stabilization buffer is prepared in advance with optimal concentrations of guanidine hydrochloride (4.05-4.95 M), TRITON X-100 (0.09-0.11% v/v), and EDTA (18-22 mM), adjusted to pH less than 4.5, and placed in collection tubes before sample arrival. This preliminary preparation ensures immediate RNA protection upon sample collection, eliminating the need for subsequent processing steps and enabling straightforward room temperature storage while maintaining RNA integrity.
2Reliability
If snap-freezing reagents and super-low temperature freezers are used, then RNA degradation is prevented, but sample damage and equipment complexity increase
Solution Approach 1:
The patent replaces the mechanical cooling system (freezers at -80°C) with a chemical stabilization system. The stabilization buffer containing guanidine hydrochloride, TRITON X-100, and EDTA at pH less than 4.5 chemically prevents RNA degradation through denaturation and chelation mechanisms, eliminating the need for super-low temperature equipment while maintaining RNA stability during storage and transport.
3Duration of action of stationary object
If freezing process is applied to preserve samples, then long-term storage is achieved, but biological sample damage occurs
Solution Approach 1:
The patent converts the potentially harmful freezing process into a beneficial chemical stabilization process. By using the stabilization buffer with specific concentrations of guanidine hydrochloride, TRITON X-100, and EDTA at pH less than 4.5, the sample can be stored at room temperature for extended periods without the mechanical stress of freezing and thawing, thereby preventing sample damage while achieving long-term storage durability.
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 stabilization buffer maintains RNA integrity, with minimal degradation, allowing for accurate RNA expression analysis even after incubation at room temperature for up to a day, facilitating diagnostic and research applications.
Implementation Method 1
guanidine hydrochloride at a concentration of about 4.05 M to about 4.95 M
Implementation Method 2
EDTA at a concentration of about 18 mM to about 22 mM
Implementation Method 3
TRITONTM X-100 at a concentration of about 0.09% to about 0.11% (v/v)
Implementation Method 4
wherein the pH of the stabilization buffers is less than about 4.5
Data Source
AI summary
The present disclosure provides a buffer comprising at least one chaotropic agent, at least one chelating agent and at least one non-ionic surfactant, wherein the buffer stabilizes a biological sample at about room temperature for at least about one day.