Solid Chaotropic Agent Nucleic Acid Isolation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating cell-free nucleic acids from bodily fluids, such as blood and plasma, face challenges with low yields and require high concentrations of chaotropes, leading to viscous solutions and excessive sample dilution, which are not compatible with existing automation equipment.
Innovation Solution
A process using a chaotrope in solid form, combined with a detergent and a buffer, along with magnetic particles, to form a reaction mixture that allows for efficient isolation of cell-free nucleic acids with minimal dilution and high chaotrope concentrations, reducing the need for carrier nucleic acids and minimizing sample volume.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If high concentrations of chaotropes are used to facilitate dissociation of cell-free nucleic acid from proteins, then nucleic acid release is improved, but the solution becomes viscous and difficult to work with
Solution Approach 1:
The patent changes the physical state parameter of the chaotrope from liquid to solid form. Solid chaotropes can be dissolved in minimal volumes of aqueous buffer to achieve high working concentrations (e.g., 5-10 M), providing effective nucleic acid release while avoiding the viscosity problems associated with liquid chaotrope solutions. This parameter change allows high chaotrope concentrations to be achieved without compromising solution handling characteristics.
2Reliability
If high molarities of chaotropes are used to release nucleic acids, then capture on solid phase is improved, but several-fold dilution of biological fluid sample is required causing undesirably high fluid volumes
Solution Approach 1:
The patent changes the concentration parameter by using solid chaotropes that can be dissolved in minimal volumes to achieve high working concentrations. This eliminates the need for several-fold dilution of the biological fluid sample, thereby maintaining small sample volumes (e.g., 50-500 μL) while still achieving effective nucleic acid capture on the solid phase.
Solution Approach 2:
The patent uses carrier nucleic acid molecules that are incorporated into the solid phase matrix. These carrier molecules serve as a template or copy that facilitates the capture and retention of target nucleic acids through hybridization or adsorption mechanisms, improving capture efficiency without requiring large sample volumes or high chaotrope dilutions.
3Reliability
If existing methods are used to purify nucleic acids from biological fluids, then purification is achieved, but yields are very low and do not work well when extracting small amounts of nucleic acids from large samples
Solution Approach 1:
The patent optimizes the concentration parameter by using solid chaotropes dissolved in minimal volumes, creating highly concentrated reaction mixtures that maximize nucleic acid release efficiency. This parameter optimization enables effective extraction of small amounts of nucleic acids from relatively small sample volumes, dramatically improving yield compared to conventional methods that require large sample volumes and produce low yields.
Solution Approach 2:
The patent incorporates carrier nucleic acids into the solid phase that serve as templates for capturing target nucleic acids. This copying mechanism enhances the efficiency of nucleic acid recovery by providing multiple binding sites and facilitating the capture of even trace amounts of target nucleic acids, thereby improving overall yield.
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 enables optimal direct capture of cell-free nucleic acids with reduced sample handling costs and increased throughput, achieving higher chaotrope concentrations with minimal fluid volumes, particularly beneficial for prenatal diagnostics.
Implementation Method 1
Some methods entail use of chaotropes which facilitate dissociation of cell-free nucleic acid from proteins
Implementation Method 2
capture on a solid phase such as a silicon phase
Implementation Method 3
magnetic particles, thus forming a reaction mixture containing the cell-free nucleic acid; magnetically separating the solid phase having the cell-free nucleic acid bound thereto from the reaction mixture
Data Source
AI summary
Disclosed is a process for isolating cell-free nucleic acid (including both DNA and RNA) or an analog thereof from a bodily fluid, and which entails: a) mixing in a container the bodily fluid, a chaotropic agent in solid form, a detergent and a buffer, and a solid phase which includes magnetic particles, thus forming a reaction mixture containing the cell-free nucleic acid; b) magnetically separating the solid phase having the cell-free nucleic acid bound thereto from the reaction mixture; and optionally c) dissociating the nucleic acid from the solid phase. Compositions and kits are also disclosed.