Nucleic Acid Purification Device with Nested Column
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Solution Overview
Problem
Current methods for isolating and purifying nucleic acids from large sample volumes face challenges such as limited sample volume capacity, high buffer requirements leading to diluted solutions, and inefficient elution processes, which hinder effective nucleic acid recovery for diagnostic applications like detecting tumor cells.
Innovation Solution
A device comprising a detachable funnel, separating column, and tab with a nucleic acid-binding material, allowing for efficient binding, washing, and elution of nucleic acids, utilizing chaotropic reagents to facilitate reversible binding to glass surfaces, enabling higher yield and purity of nucleic acids from larger sample volumes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If large sample volumes are processed using conventional methods, then nucleic acid recovery is achieved, but the sample volume capacity is limited and buffer requirements lead to diluted solutions
Solution Approach 1:
The device employs a nested structure where an inner column containing nucleic acid-binding material is placed inside an outer collection vessel. This allows the inner column to be processed independently while the outer vessel collects filtrate and washes, enabling efficient handling of large sample volumes without requiring multiple separate devices.
Solution Approach 2:
The device is divided into functionally independent segments: an inner column for nucleic acid binding, an outer collection vessel for filtrate collection, and a tab for support. This segmentation allows each component to be optimized for its specific function and facilitates easy assembly, disassembly, and sterilization.
2Quantity of substance
If conventional binding methods are used, then nucleic acids bind to glass surfaces, but large amounts of buffer are necessary for effective elution resulting in diluted solutions
Solution Approach 1:
The device utilizes chaotropic salts to fundamentally change the binding parameters, allowing nucleic acids to bind irreversibly to the nucleic acid-binding material under chaotropic conditions. This enables elution with small volumes of buffer by simply removing the chaotropic agent, avoiding the need for large amounts of elution buffer and preventing dilution of the final nucleic acid solution.
3Productivity
If multiple devices in strip and 96-well microwell plate format are used, then processing speed and throughput are improved, but the volume of samples is often limited
Solution Approach 1:
The nested configuration of the inner column within the outer collection vessel allows the device to accommodate large sample volumes while maintaining a compact footprint suitable for high-throughput processing. Multiple such devices can be processed simultaneously in standard centrifuges or vacuum manifolds, achieving both high throughput and large sample volume capacity.
4Reliability
If glass fleeces or silica gel suspensions are used for batch process, then nucleic acids are bound and separated, but additional preparation steps are required for certain applications due to diluted solutions
Solution Approach 1:
By using chaotropic salts to establish irreversible binding conditions, the device eliminates the need for additional concentration or purification steps. The nucleic acids are bound under chaotropic conditions and can be eluted directly in small volumes of buffer, providing both high purity and readiness for downstream applications without time-consuming additional preparation.
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 device enhances nucleic acid yield and purity by allowing efficient binding and elution processes, particularly from larger volumes, improving the sensitivity and throughput of nucleic acid isolation for diagnostic applications.
Implementation Method 1
utilize the property of nucleic acids that has been known since the end of the seventies of binding under chaotropic salt conditions to glass surfaces
Implementation Method 2
Centrifugation vessels with inserted glass fleeces or silica gel suspensions which allow a batch process
Implementation Method 3
utilizing chaotropic reagents to facilitate reversible binding to glass surfaces, enabling higher yield and purity of nucleic acids
Data Source
AI summary
The invention concerns a method for isolating and purifying nucleic acids from large sample volumes and a device suitable for this. The device comprises a funnel, a separating column, and a tab which are detachable from each other. The method according to the invention in which the device according to the invention is used ensures a higher yield of nucleic acids.