Low-Speed Centrifugation for Nucleic Acid-Containing Particle Isolation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for isolating nucleic acid-containing particles from biological samples, such as exosomes and microvesicles, are inefficient and do not yield high-quality nucleic acids suitable for accurate analysis, particularly due to RNA degradation and other adverse factors.
Innovation Solution
A method involving low-speed centrifugation (up to 200,000 g) is used to pellet particles, followed by treatment with RNase inhibitors and other agents to mitigate adverse factors, enabling extraction of high-quality nucleic acids, including 18S and 28S rRNA, from these particles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high-speed centrifugation is used to isolate microvesicles, then isolation efficiency is improved, but nucleic acid quality deteriorates due to degradation
Solution Approach 1:
The patent changes the centrifugation speed parameter from high-speed to low-speed (below 200,000 g) to resolve the contradiction. This parameter change allows sufficient isolation of microvesicles while preventing the mechanical stress that causes nucleic acid degradation, thereby maintaining both isolation efficiency and nucleic acid quality
Solution Approach 2:
The patent applies preliminary protection measures by adding RNase inhibitors and other protective agents to the isolation buffer before centrifugation. This preliminary action prevents nucleic acid degradation from occurring during the isolation process, allowing efficient isolation without quality loss
2Manufacturing precision
If extensive purification steps are used to remove contaminants, then purity is improved, but loss of substance increases
Solution Approach 1:
The patent selectively extracts only the necessary protective components (RNase inhibitors, protective agents) from the complex mixture of purification reagents. This targeted extraction approach removes contaminants while preserving nucleic acids, achieving high purity without excessive loss of substance
Solution Approach 2:
The patent uses protective agents as intermediaries that bind to potential contaminants or degradation enzymes, allowing their removal without directly exposing nucleic acids to harsh purification conditions. This intermediary approach maintains nucleic acid integrity and yield while achieving purification
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 method achieves high-quality nucleic acid extraction with intact ribosomal RNA ratios and integrity, suitable for downstream analysis, including gene expression and mRNA analysis, from biological samples like serum and urine.
Implementation Method 1
isolating nucleic acid-containing particles from a biological sample by one or more centrifugation procedures at a speed not exceeding about 200,000 g
Data Source
AI summary
A method for extracting nucleic acids from a biological sample by isolating nucleic acid-containing particles from the biological sample by one or more centrifugation procedures, performing one or more steps to mitigate adverse factors that prevent or might prevent high quality nucleic acid extraction, and extracting nucleic acids from the isolated particles. The centrifugation procedures are performed at a speed not exceeding about 200,000 g. The extracted nucleic acids contain both 18S and 28S rRNA.