Nucleic Acid Extraction Using High-Temperature Chemical Lysis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nucleic acid extraction methods, particularly those using enzymatic lysis, are time-consuming and require specific kits or protocols for different biological materials, making them unsuitable for applications needing promptness.
Innovation Solution
A method involving mixing a biological sample with lithium dodecyl sulfate and a chaotropic compound, followed by the addition of 1-propanol and silica particles at elevated temperatures to adsorb nucleic acids, which are then washed and eluted, reducing the overall extraction time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If enzymatic lysis method is used to extract nucleic acid, then nucleic acid can be effectively extracted, but the extraction time becomes excessively long
Solution Approach 1:
The patent changes the temperature parameter from conventional low temperatures to high temperature (95-100°C) to accelerate the lysis process. By performing lysis at high temperature, the extraction time is significantly reduced while maintaining effective nucleic acid release from cells
Solution Approach 2:
The patent replaces enzymatic lysis with a chemical lysis system using SDS (anionic surfactant) and chaotropic compounds. This substitution eliminates the time-consuming enzymatic digestion process while achieving effective cell membrane disruption and nucleic acid release through chemical mechanisms
2Reliability
If different kits or protocols are used for different biological materials, then extraction effectiveness is optimized, but device complexity and operational difficulty increase
Solution Approach 1:
The patent develops a universal extraction kit and protocol that can effectively extract nucleic acids from diverse biological materials including bacteria, fungi, viruses, and mammalian cells. The high-temperature chemical lysis system with SDS and chaotropic compounds provides a single standardized procedure that works across multiple sample types, eliminating the need for multiple specialized kits
3Reliability
If centrifugal separation step is added to the extraction process, then nucleic acid purification is improved, but the extraction time increases
Solution Approach 1:
The patent extracts and removes contaminants through the chemical lysis and precipitation process itself, where nucleic acids are selectively precipitated while contaminants remain in solution. This eliminates the need for separate centrifugal separation steps, achieving both purification and time reduction
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 significantly reduces the time required for nucleic acid extraction from biological samples, allowing for high-yield extraction across various biological materials with a single kit and protocol.
Implementation Method 1
mixing the lysate, an alkanol having a boiling point higher than 75°C, namely 1-propanol, a chaotropic compound and silica particles to adsorb the nucleic acid on the silica particles
Implementation Method 2
a chaotropic compound, which is a substance that robs hydration water from biopolymers such as nucleic acid, and destabilize the higher-order structure of the biopolymers
Implementation Method 3
mixing a biological sample, an anionic surfactant, namely lithium dodecyl sulfate (LDS), and a chaotropic compound to obtain a lysate
Data Source
Figure 1(a)~1(e)
Figure 2(a)~2(b)
Figure 3(a)~3(b)
AI summary
A method for extracting nucleic acid from a biological sample is disclosed. The method comprises steps of: (i) mixing a biological sample, an anionic surfactant and a chaotropic compound to obtain a lysate; (ii) mixing the lysate, an alkanol having a boiling point higher than 75°C, a chaotropic compound and silica particles to adsorb the nucleic acid on the silica particles; (iii) washing the silica particles having the nucleic acid adsorbed thereon with a washing liquid; and (iv) eluting the nucleic acid adsorbed on the silica particles with an eluent, wherein at least steps (i) and (ii) are performed at a temperature higher than 75°C.