Polyol-Based Nucleic Acid Isolation Method
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Solution Overview
Problem
Current methods for nucleic acid isolation from biological samples are cumbersome, time-consuming, and often involve the use of volatile solvents like ethanol and isopropanol, which can interfere with downstream processes and increase costs due to flammability and handling issues.
Innovation Solution
The use of polyols, such as undiluted or aqueous compositions of C2-C10 alcohols with at least two hydroxy groups, to precipitate or maintain nucleic acids out of solution, replacing traditional solvents and reducing viscosity, thereby facilitating efficient nucleic acid isolation and amplification without the need for alcohol-based wash buffers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If ethanol or isopropanol is used for nucleic acid precipitation and washing, then nucleic acid can be effectively isolated from biological samples, but the process becomes time-consuming and requires multiple complex steps
Solution Approach 1:
The patent changes the chemical parameter of the precipitation agent from monohydric alcohols (ethanol, isopropanol) to polyols (glycerol, ethylene glycol, propylene glycol). This parameter change allows nucleic acid isolation to be achieved in a single step without requiring multiple washing steps, thereby reducing processing time while maintaining isolation efficiency
Solution Approach 2:
The patent combines the precipitation and washing functions into a single step by using polyols that can both precipitate nucleic acid and serve as the washing medium. This merging of steps eliminates the need for separate washing operations, reducing both time and procedural complexity
2Manufacturing precision
If multiple extraction and washing steps are performed, then nucleic acid purity is improved, but the process becomes more complex and laborious
Solution Approach 1:
The patent merges multiple extraction and washing steps into a single operation by using polyols that simultaneously achieve precipitation, washing, and contamination removal. This single-step process maintains nucleic acid purity while eliminating the complexity of multiple sequential steps
Solution Approach 2:
The polyol reagent performs multiple functions simultaneously: it precipitates nucleic acid, washes the precipitate, removes contaminants, and prevents degradation. This multi-functionality replaces what traditionally required multiple specialized reagents and steps, simplifying the overall process while maintaining purity
3Productivity
If volatile solvents like ethanol and isopropanol are used, then nucleic acid precipitation is effective, but handling costs increase due to flammability and interference with downstream processes
Solution Approach 1:
The patent changes the chemical properties of the precipitation agent by replacing volatile, flammable monohydric alcohols with non-volatile, non-flammable polyols. This parameter change eliminates safety hazards and removes interference with downstream enzymatic reactions while maintaining precipitation efficiency
Solution Approach 2:
The patent converts the potential harm of using alcohol-based reagents (flammability, interference with enzymes) into a benefit by selecting polyols that are inherently safer and compatible with downstream processes. The polyols provide the same precipitation function without the harmful side effects
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 approach allows for rapid and efficient nucleic acid isolation with minimal sample loss and contamination, maintaining high amplification efficiency in processes like qPCR, while eliminating the use of volatile solvents and reducing operational costs.
Implementation Method 1
use of a polyol to cause a nucleic acid in solution in an aqueous medium to leave the solution phase by combining the aqueous medium with the polyol
Implementation Method 2
polyols are not solvents for nucleic acids and have 'anti-solvating' activity in relation to nucleic acids
Data Source
AI summary
A method for processing a nucleic acid, in which the nucleic acid is exposed to an aqueous medium which includes a polyol in sufficient proportion for at least a portion of the nucleic acid to enter or remain in an extra-solution phase. Thus, a polyol may be used to bind a nucleic acid which is in solution to a solid support or to wash a nucleic acid on a solid support whilst maintaining it on the support. The polyol may for example be a C2-C10 alkanediol.