RNA Extraction Reagent Using Heparin and Reducing Agents
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Solution Overview
Problem
Current RNA extraction methods from eukaryotic cells are harsh, require hazardous chemicals, and involve multiple steps, including the use of toxic organic solvents like chloroform, which poses environmental concerns and is time-consuming.
Innovation Solution
A reagent comprising heparin, a reducing agent, a chelating agent, a buffer, and an alkali metal halide, optionally with a nonionic detergent, that disrupts RNA structures and proteins without using organic solvents, allowing for efficient RNA extraction in a short time with minimal chemical and environmental impact.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional RNA extraction reagents (containing ionic detergents, chelators, reducing agents) are used, then RNA extraction effectiveness is improved, but the reagent becomes harsh and requires toxic organic solvents like chloroform
Solution Approach 1:
The patent changes the chemical parameters of the extraction reagent by replacing ionic detergents with nonionic detergents and eliminating the need for toxic organic solvents like chloroform. The reagent formulation includes specific concentrations of nonionic detergent (0.1-5%), chelator (0.1-5 mM), reducing agent (0.1-5 mM), and heparin (1-100 μg/mL), creating a non-toxic alternative that maintains extraction effectiveness while removing harmful components
Solution Approach 2:
The patent extracts and removes the harmful components (ionic detergents, chloroform, and other toxic chemicals) from the traditional extraction reagent formulation while retaining the essential functional components needed for RNA extraction. This results in a purified reagent system that achieves the same extraction goals without the associated toxicity and environmental harm
2Reliability
If traditional RNA extraction procedures are used, then RNA can be extracted, but the process requires multiple steps and takes several hours
Solution Approach 1:
The patent merges multiple extraction steps into a single integrated reagent system that performs cell lysis, RNA stabilization, and protein disruption simultaneously. The combined reagent formulation contains all necessary components (nonionic detergent, chelator, reducing agent, heparin) that work together in one step to achieve RNA extraction, eliminating the need for sequential procedures and reducing total extraction time from several hours to minutes
3Productivity
If harsh reagents and organic solvents are used for RNA extraction, then extraction efficiency is improved, but safety for users and environmental impact deteriorate
Solution Approach 1:
The patent employs a disposable, single-use reagent formulation that is safe for users and environmentally benign. The reagent contains no toxic components, allowing it to be discarded after use without special handling or disposal procedures. This approach maintains high extraction efficiency while eliminating safety concerns and environmental harm associated with traditional harsh reagents and organic solvents
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 reagent effectively extracts RNA from eukaryotic cells in a few steps and within minutes, maintaining RNA integrity without disrupting cellular material or the environment, and is safer for users.
Implementation Method 1
The reagent contains from 1 pg/μl to 10 pg/μl heparin
Implementation Method 2
from 1 mM to 10 mM of a reducing agent effective to reduce disulfide bonds in cellular probes
Implementation Method 3
a chelating agent in an amount sufficient to disrupt RNA secondary and tertiary structures
Data Source
AI summary
RNA is extracted from cellular material with a reagent that includes heparin, a reducing agent to reduce disulfide bonds, a chelating agent, a buffer, and an alkali metal halide. The reagent does not require the use of organic solvents, and the reagent allows extraction to be performed in a relatively short period of time in comparison to the prior art.