RNA Binding Buffer Using High Flash Point Solvents
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Solution Overview
Problem
Current RNA isolation methods using chaotropic agents and organic solvents face issues with RNA yield, integrity, and DNA contamination, and are often hazardous due to flammability and toxicity concerns.
Innovation Solution
A phenol-free RNA binding buffer containing a chaotropic agent and organic solvents like ethylene carbonate, ethylene glycol diacetate, or 2-pyrrolidone is used to selectively bind RNA to a solid phase, enhancing yield and reducing DNA contamination while being safer and more cost-effective.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional organic solvents (methanol, ethanol, acetone) are used in RNA binding buffer, then RNA binding efficiency is achieved, but flash point is low and fire risk increases
Solution Approach 1:
The patent changes the chemical parameters of the organic solvent by selecting compounds with high flash points (ethylene carbonate: 124°C, ethylene glycol diacetate: 170°C, 2-pyrrolidone: 207°C) while maintaining the functional properties needed for RNA binding. This parameter substitution resolves the contradiction by achieving the same binding efficiency without the fire hazard of traditional solvents like ethanol (flash point: 13°C) or acetone (flash point: -20°C).
2Reliability
If traditional organic solvents are used in RNA binding buffer, then RNA binding is achieved, but toxicity is high and user safety decreases
Solution Approach 1:
The patent changes the toxicity parameter by selecting organic solvents with favorable safety profiles. Ethylene carbonate, ethylene glycol diacetate, and 2-pyrrolidone are chosen because they exhibit lower acute toxicity compared to traditional solvents like phenol, chloroform, or high concentrations of ethanol, thereby improving user safety while maintaining RNA binding capability.
3Reliability
If traditional organic solvents are used in RNA binding buffer, then RNA binding occurs, but RNA yield and integrity are suboptimal
Solution Approach 1:
The patent optimizes the concentration parameters of the organic solvent in the binding buffer to achieve superior RNA recovery. By carefully controlling the amount of ethylene carbonate, ethylene glycol diacetate, or 2-pyrrolidone added to the chaotropic lysis buffer, the invention maximizes RNA binding efficiency and minimizes losses, thereby improving both yield and integrity compared to traditional solvent systems.
4Reliability
If traditional organic solvents are used in RNA binding buffer, then RNA binding is achieved, but DNA contamination remains critical
Solution Approach 1:
The patent applies local quality differentiation by creating distinct chemical environments that favor selective RNA binding. The high flash point organic solvents interact differently with RNA versus DNA in the chaotropic buffer system, creating local chemical conditions that enhance RNA-solvent binding affinity while reducing DNA solubility, thereby improving selectivity and reducing DNA contamination in the final RNA preparation.
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 solution significantly increases RNA yield and integrity, reduces DNA contamination, and is safer and more cost-effective, with RNA integrity numbers often above 9.0, and can be used with a broad spectrum of biological samples.
Implementation Method 1
the moderate solubility of the claimed organic solvents in water is substantially increased by the addition of the chaotropic agent to the buffer
Implementation Method 2
allows RNA to bind selectively to an RNA-binding solid phase
Data Source
AI summary
The subject innovation relates to a RNA binding buffer comprising (a) at least one chaotropic agent; and (b) an organic solvent selected from the group consisting of ethylene carbonate, ethylene glycol diacetate and 2-pyrrolidone or combinations thereof. The subject innovation further relates to a method of binding RNA to a solid support and to a method of isolating RNA both making use of the binding buffer of the subject innovation. The subject innovation finally relates to a kit comprising the RNA binding buffer or the organic solvent as relevant substance therein.