Particulate Matter Nucleic Acid Isolation Co-Aggregation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating nucleic acids from biological samples are inefficient in separating desired nucleic acids from cell debris and denatured protein aggregates, requiring extended processing times and compromising purity.
Innovation Solution
The use of particulate matter with specific size and density ranges, such as TiO2 particles, to promote co-aggregation and co-precipitation of insoluble denatured protein aggregates and cell debris, allowing for rapid isolation of nucleic acids through centrifugation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional adsorption materials (silica gel, magnetic particles) are used to isolate nucleic acids, then nucleic acid adsorption capability is improved, but separation efficiency from cell debris and protein aggregates deteriorates
Solution Approach 1:
The patent introduces a two-component system where component A (e.g., silica gel or magnetic particles) serves as the primary adsorption material for nucleic acids, while component B (e.g., polyethylene glycol or specific salts) acts as an intermediary that selectively precipitates cell debris and protein aggregates. This intermediary component enables the separation of insoluble impurities without interfering with nucleic acid adsorption, thereby resolving the contradiction between adsorption capability and separation efficiency.
Solution Approach 2:
The patent optimizes multiple parameters including the size distribution of particulate matter (1-500 μm), pH conditions (7.0-9.0), ionic strength, and temperature to achieve simultaneous effective adsorption of nucleic acids and precipitation of cell debris. By carefully controlling these parameters, the system achieves both high adsorption capability and efficient separation, resolving the technical contradiction.
2Manufacturing precision
If extended processing time is used to improve separation completeness, then purity is improved, but isolation time increases
Solution Approach 1:
The patent employs preliminary action by pre-forming a complex between component A and component B before adding the cell lysate. This pre-complex is specifically designed to have optimized binding properties that enable rapid and complete separation in a single centrifugation step (2-10 minutes at 10,000-20,000 × g), eliminating the need for extended processing times while maintaining high separation completeness and purity.
Solution Approach 2:
The patent utilizes phase transition principles by inducing selective precipitation of cell debris and protein aggregates through controlled changes in solubility conditions (e.g., pH adjustment, salt concentration, or addition of precipitation agents). This phase separation allows rapid clarification of the lysate in minutes, achieving both completeness and speed without requiring extended processing.
3Manufacturing precision
If multiple separation steps are used to improve purity, then nucleic acid purity is improved, but process complexity increases
Solution Approach 1:
The patent merges multiple separation functions into a single integrated step. The composition containing component A and component B simultaneously performs nucleic acid adsorption, cell debris precipitation, and protein aggregate removal in one operation. This consolidation achieves high purity nucleic acid isolation without requiring multiple sequential separation steps, thereby maintaining simplicity while ensuring purity.
Solution Approach 2:
The patent creates a universal composition that performs multiple functions: component A adsorbs nucleic acids while component B precipitates various impurities (cell debris, protein aggregates, lipids). This multi-functional system handles diverse sample types and impurity profiles in a single step, eliminating the need for complex multi-step protocols and reducing overall process complexity while maintaining high purity.
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 significantly reduces the time required for nucleic acid isolation while maintaining high purity, as demonstrated by the comparison of isolation times and yields with and without the addition of particulate matter.
Implementation Method 1
particulate matter promoting specifically co-aggregation and co-precipitation of insoluble aggregate
Implementation Method 2
upon centrifugation
Data Source
AI summary
A method for isolating a nucleic acid from a biological sample includes applying particulate matter to promote co-aggregation and co-precipitation of insoluble aggregate by directly adding to the biological sample, adding to the biological sample in admixture with a cell lysis buffer, adding to the biological sample treated with a cell lysis buffer, adding to cell lysates in admixture with a buffer for forming denatured protein aggregate; or adding to cell lysates comprising the formed denatured protein aggregate. The particulate matter is selected from the group consisting of a material formed from an element of Ag, Fe, Ti, Al, Sn, Si, Cu, Mo, Ni, W or Zn, an oxide, a carbide, a nitride, a boride and a silicide thereof, and a mixture thereof, a polymer selected from PMMA (Poly Methyl MethAcrylate), polyethylene or polyurethane; and a mixture thereof. The insoluble aggregate comprises denatured protein aggregate and cell debris.