Nucleic Acid Extraction via High-Temperature Sealed Vessel Lysis
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Solution Overview
Problem
Current nucleic acid extraction methods, particularly enzymatic or chemical lysis methods, are time-consuming and require multiple steps, making them unsuitable for rapid detection applications, and lack a uniform protocol for different cell types, complicating the process, especially for unknown microbe testing.
Innovation Solution
A method involving heating a fungal cell suspension to a temperature range of 125°C to 160°C in a sealed vessel, with the option to blend a cell lysis accelerator before or after heating, to efficiently extract nucleic acids from fungal cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If enzymatic or chemical lysis methods are used for nucleic acid extraction, then nucleic acid can be extracted from cells, but the treatment time becomes excessively long and the process becomes complicated
Solution Approach 1:
The invention changes the temperature parameter dramatically by heating the lysis mixture to 100°C or higher, which fundamentally alters the lysis mechanism from slow enzymatic/chemical reactions to rapid thermal denaturation and cell wall disruption, reducing lysis time from hours to minutes while maintaining extraction effectiveness
Solution Approach 2:
The invention employs periodic heating cycles where the lysis mixture is heated to high temperature for a short duration and then cooled, repeating this cycle multiple times. This periodic thermal action progressively disrupts cell structures and releases nucleic acids efficiently without requiring prolonged continuous heating
2Adaptability or versatility
If different enzymes or chemical protocols are used for different cell types, then specific cell lysis can be achieved, but the protocol complexity increases
Solution Approach 1:
The invention creates a universal lysis protocol that works across diverse cell types (bacteria, fungi, plant cells, animal cells) by using high-temperature heating that physically disrupts all types of cell walls and membranes simultaneously, eliminating the need to select different enzymes or chemicals for different organisms
Solution Approach 2:
The invention replaces the complex biochemical system (multiple enzymes and chemicals tailored to specific cell types) with a simple physical system (high-temperature heating), which universally disrupts all cell structures through thermal energy without requiring knowledge of or adaptation to specific cell wall compositions
3Manufacturing precision
If long enzyme reaction time is used to obtain long-chain nucleic acid, then nucleic acid quality is improved, but the method lacks rapidity
Solution Approach 1:
The periodic heating and cooling cycles allow nucleic acids to be released and protected in a controlled manner, with cooling periods preventing excessive degradation while heating periods ensure complete cell disruption, achieving both long-chain integrity and rapid processing
Solution Approach 2:
The invention rushes through the lysis process by applying high temperature briefly to achieve complete cell disruption and nucleic acid release in minutes rather than hours, then immediately cools to preserve the released nucleic acids, skipping the prolonged gradual lysis required by traditional methods
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 cell lysis and nucleic acid extraction, achieving high efficiency and adaptability across various fungal cell types, while maintaining DNA integrity for PCR amplification.
Implementation Method 1
heating the fungal cell suspension housed in the vessel up to a prescribed highest temperature with the vessel held in a sealed state, wherein the prescribed highest temperature is in a range of 125°C to 160°C
Data Source
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AI summary
There is provided a nucleic acid extraction method applicable to microbes in a relatively wide range, and capable of rapidly extracting nucleic acid. The nucleic acid extraction method comprises the steps of introducing a cell suspension into a vessel, sealing the vessel, and preheating a heater up to a set temperature not lower than 100°C. Further, the method comprises the step of bringing the vessel into contact with the heater heated up to the set temperature, thereby heating the cell suspension housed in the vessel up to a prescribed highest temperature at not lower than 100°C with the vessel held in a sealed state.