Small RNA Purification via Bicarbonate Extraction
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Solution Overview
Problem
Existing methods for isolating small RNAs from fungi and plants are not suitable for industrial-scale use due to the need for toxic solvents, expensive reagents, and cumbersome processes, and often require solid supports for extraction, which complicates the process and increases costs.
Innovation Solution
A method involving the use of a bicarbonate solution to extract small RNAs without the need for solid supports, involving steps such as incubation, alcohol precipitation, and centrifugation, which simplifies and cost-reduces the process while maintaining yield and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If conventional methods using toxic solvents and solid supports are used, then small RNA isolation can be achieved, but the process becomes complex, expensive, and unsuitable for industrial scale-up
Solution Approach 1:
The invention extracts and removes the harmful elements (toxic solvents and solid supports) from the conventional isolation process, retaining only the essential extraction function through a simplified aqueous bicarbonate solution method that achieves the same isolation effectiveness without the complicating factors
Solution Approach 2:
The invention changes the chemical parameters of the extraction medium from toxic organic solvents and chaotropic agents to a safe aqueous bicarbonate solution, fundamentally altering the extraction chemistry to eliminate harmful substances while maintaining isolation effectiveness
2Manufacturing precision
If solid supports are used for RNA binding, then selective extraction of small RNAs is achieved, but the process requires additional steps and equipment that complicate manufacturing
Solution Approach 1:
The invention removes the solid support component entirely from the extraction process, achieving selective small RNA extraction through the chemical properties of the aqueous bicarbonate solution alone, thereby eliminating the need for filtration, centrifugation, and other solid support-related steps
Solution Approach 2:
The aqueous bicarbonate solution performs multiple functions simultaneously - it extracts small RNAs selectively, maintains RNA stability, and enables straightforward separation - without requiring the assistance of solid supports or additional processing equipment
3Reliability
If toxic solvents and expensive reagents are used, then RNA protection from degradation is achieved, but the cost and safety issues prevent industrial application
Solution Approach 1:
The invention converts the traditionally harmful aqueous environment (which would normally promote RNase activity and RNA degradation) into a beneficial protective medium by using bicarbonate solution to create conditions that inhibit RNase activity without requiring toxic solvents or chaotropic agents
Solution Approach 2:
The invention replaces expensive, toxic reagents with a cheap, safe, and readily available aqueous bicarbonate solution that can be used in large quantities for industrial-scale extraction without safety concerns or disposal issues
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 effectively isolates small RNAs with high yield and purity, eliminating the need for toxic solvents and solid supports, making it more efficient and scalable for industrial applications.
Implementation Method 1
at least one step of treating said fungi or plant or part thereof with a bicarbonate solution, to obtain a liquid phase comprising or consisting of a small RNA containing extract
Implementation Method 2
adding an alcohol solution to said liquid phase of step a) or to said concentrated small RNAs containing extract of step b') to allow precipitation of the small RNAs containing extract
Implementation Method 3
involving steps such as incubation, alcohol precipitation, and centrifugation
Data Source
AI summary
The present invention refers to a method for obtaining a small RNAs (sRNAs) containing extract from fungi and/or plant or part thereof, said extract being characterized by a fraction enriched of sRNA molecules said sRNA molecules having less than 200 nucleotides or base pairs (bp), wherein said process comprises at least one step of treating said fungi or plant or part thereof with a bicarbonate solution, to obtain a liquid phase comprising or consisting of a small RNA containing extract.