Porous Membrane RNA Purification Cartridge
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Solution Overview
Problem
Current methods for separating and purifying RNA from DNA in mixed nucleic acid samples are inefficient, laborious, and costly, with existing technologies lacking in automation, miniaturization, and requiring excessive DNase for selective RNA recovery.
Innovation Solution
A method using a cartridge with a nucleic acid-adsorbing porous membrane, where nucleic acid is adsorbed, washed, subjected to DNase treatment, and RNA is desorbed using a recovering solution, with a DNase solution amount of 130 μl or less per 1 cm² and a concentration of 10 to 10,000 Kunitz U/mL, enabling efficient and cost-effective RNA purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If nucleic acid is adsorbed to silicon dioxide, silica polymer, magnesium silicate or other solid phase for separation and purification, then separation performance is improved, but automation aptitude deteriorates and device complexity increases
Solution Approach 1:
The patent employs disposable cartridges containing pre-loaded porous membranes with adsorbed DNase. These single-use cartridges eliminate the need for complex automation while maintaining separation performance, as each cartridge is designed for one-time use and then discarded, avoiding the need for sterilization and reuse protocols that would complicate automation.
Solution Approach 2:
The system divides the purification process into discrete functional modules within the cartridge: sample loading zone, DNase treatment zone with porous membrane, washing zone, and elution zone. This segmentation allows manual operation while maintaining separation performance, and the modular design could facilitate future automation if needed.
2Reliability
If a porous membrane with sufficient thickness is used to obtain mechanical strength for homogeneous DNase interaction, then reliability is improved, but DNase solution quantity increases and cost rises
Solution Approach 1:
The patent uses porous membranes with controlled pore structures that allow DNase to penetrate and interact homogeneously throughout the membrane matrix. The porosity enables enzymatic action throughout the membrane volume rather than just at the surface, maintaining reliability with thinner membranes and reducing DNase solution requirements.
Solution Approach 2:
The patent optimizes membrane thickness parameters and DNase concentration parameters to achieve the minimum necessary thickness for mechanical strength while maximizing DNase penetration efficiency. By changing these parameters within specific ranges, the system maintains reliability without requiring excessive DNase solution.
3Manufacturing precision
If multiple washing and desorption steps are performed to achieve high-purity RNA separation, then purity is improved, but loss of time increases and productivity decreases
Solution Approach 1:
The cartridge is pre-conditioned during manufacturing with optimized washing and desorption reagents positioned in specific zones. This preliminary preparation allows the user to achieve high-purity RNA separation by simply passing solutions through the pre-programmed cartridge, eliminating the need for multiple manual washing and desorption steps while maintaining productivity.
Solution Approach 2:
The patent combines multiple functions into single integrated steps: the cartridge design allows washing and desorption to occur in a single continuous flow through different zones of the membrane, merging what would traditionally be separate操作步骤 into one streamlined process that maintains purity while improving speed.
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 achieves high-purity RNA separation with improved automation, miniaturization, and reduced DNase usage, allowing for rapid and efficient RNA recovery with minimal impurities and operational costs.
Implementation Method 1
nucleic acid is adsorbed to silicon dioxide, silica polymer, magnesium silicate or the like solid phase
Implementation Method 2
a porous membrane which the solutions can pass through
Implementation Method 3
degrading DNA with DNase for selectively recovering RNA from a mixture sample containing DNA and RNA
Data Source
AI summary
A method for selectively separating and purifying RNA from a mixture solution of nucleic acid containing DNA and RNA, wherein the method comprising the steps of: (1-a) adsorbing nucleic acid; (1-b) washing; (1-c) subjecting to a DNase treatment; (1-d) washing; and (1-e) desorbing the RNA from a nucleic acid-adsorbing porous membrane by a recovering solution, wherein in the step (1-c), a total amount of a DNase solution is 130 μl or less per 1 cm2 of the membrane. And a method for selectively separating and purifying RNA or DNA, which comprises the steps of: (2-a) adsorbing nucleic acid; (2-b) washing by a washing solution; and (2-c) desorbing the nucleic acid from a nucleic acid-adsorbing porous membrane, wherein the washing solution contains a water-soluble organic solvent having a concentration of 50% by weight or less, and does not contain a chaotropic salt.