Magnetic Nucleic Acid Purification Apparatus
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Solution Overview
Problem
Current methods for isolating DNA and RNA from biological samples are often inefficient and require multiple steps, involving hazardous reagents and large, expensive instruments, with RNA being particularly sensitive to degradation.
Innovation Solution
An automated method using lithium salts, surfactants, and magnetic particles to lyse cells and bind nucleic acids, followed by magnetic separation, which reduces the need for chaotropic salts and phenol/chloroform, and allows for simultaneous isolation of DNA and RNA from the same sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional methods using glass fiber columns and chaotropic salts are used, then nucleic acid purification can be achieved, but the process becomes complex and requires large, expensive instruments
Solution Approach 1:
The patent replaces complex mechanical filtration systems (glass fiber columns) with a magnetic field-based separation system. Magnetic particles functionalized with nucleic acid-binding groups allow purification through simple magnetic separation, eliminating the need for large column-based instruments and reducing device complexity while maintaining purification reliability
Solution Approach 2:
The patent changes the chemical parameters by replacing chaotropic salts (guanidine) with lithium salts and alcohol-based solutions. This parameter change enables effective nucleic acid purification without requiring the complex instrument systems needed for traditional chaotropic salt methods, thereby reducing device complexity
2Reliability
If phenol/chloroform methods are used for nucleic acid isolation, then purification can be achieved, but hazardous reagents are required
Solution Approach 1:
The patent fundamentally changes the chemical composition parameters by replacing phenol/chloroform hazardous reagents with a safe aqueous system comprising lithium salts and alcohol-based solutions. This parameter change maintains effective nucleic acid purification while completely eliminating the harmful factors associated with phenol and chloroform exposure
Solution Approach 2:
The patent employs disposable magnetic particles that can be discarded after a single use, eliminating the need for repeated sterilization and handling of hazardous reagents. This approach maintains purification reliability while removing the need for hazardous chemical agents
3Reliability
If multiple stepwise isolation processes are used for DNA and RNA, then purification can be achieved, but the process time increases
Solution Approach 1:
The patent creates a universal magnetic particle system that can purify both DNA and RNA using the same basic protocol. The magnetic particles are functionalized to bind nucleic acids generally, allowing a single multi-functional approach to replace separate stepwise processes for DNA and RNA isolation, thereby reducing total process time while maintaining purification quality
Solution Approach 2:
The patent performs preliminary cell lysis and nucleic acid release in a single step using lithium salt and alcohol-based lysis buffer, before proceeding to magnetic separation. This preliminary action consolidates multiple preparatory steps into one operation, reducing the overall time required while ensuring effective purification of both DNA and RNA
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 provides a cost-effective, efficient, and sensitive approach to isolate high-quality DNA and RNA, minimizing enzyme degradation and reducing the complexity and cost of the isolation process.
Implementation Method 1
The third step involves applying a magnetic force to the magnetic processing tube, wherein the DNA is liberated from the sample
Implementation Method 2
the second reagent contains (1) a lithium salt at a concentration of at least 9 M; (2) a surfactant concentration of at least 5%, and (3) a buffer
Implementation Method 3
The first step involves mixing a first reagent (a Lysis reagent) having a pH of at least 9, a sample, and magnetic particles to lyse the cells in the sample and thus release nucleic acid from the sample
Data Source
Figure 1A
Figure 1B
Figure 1C
AI summary
The invention relates to apparatus, materials and methods for isolating RNA or DNA from a sample . The apparatus comprises magnetic elevator means for separating magnetic particles. Aspirating and dispensing pipettes are provided, and an automated isolation method involving washing, elution, and lysis is disclosed. A lysis solution for lysing cells comprises an alkali metal salt at a concentration of about 2 M to about 10 M, a detergent at a concentration of 0.05 to 2.0% w/v, a surfactant at a concentration of about 5 to 40% w/v, a chelating agent at a concentration of about 5 to 20 mM, an antifoaming agent, a buffer, and antifoam agent of about 0.005% to about 0.1%, wherein the solution has a pH of at least 7.