Nucleic Acid Analyzer Trapezoidal Temperature Control
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Solution Overview
Problem
Existing nucleic acid analyzers face issues with overheating during PCR cycles, leading to enzyme deactivation and air bubble generation due to uniform overshooting and temperature control delays, which affect analytical performance and require fixed overshoot settings for all analysis items.
Innovation Solution
A nucleic acid analyzer with a temperature control unit that employs trapezoidal temperature control curves and adjustable overshoot settings for each PCR cycle, allowing for customizable temperature change speeds and preventing overheating by using a holder, temperature controller, and sensor to manage the reaction mixture's temperature effectively.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If uniform overshooting is performed in all PCR cycles, then the temperature of the reaction mixture reaches the target temperature faster, but the reaction mixture overheats causing enzyme deactivation and air bubble generation
Solution Approach 1:
The patent applies parameter changes by adjusting the overshoot temperature and duration based on different PCR cycle stages and reagent characteristics. Specifically, the overshoot temperature is set to 95°C for denaturation steps but reduced to 90°C or lower for extension steps, and the overshoot duration is adjusted according to the thermal stability of the nucleic acid amplification enzyme. This dynamic parameter adjustment prevents enzyme deactivation and air bubble generation while maintaining fast temperature response.
2Speed
If the temperature control block temperature is raised to overshoot the target temperature, then the reaction mixture reaches the target temperature faster, but the temperature control becomes fixed and cannot adapt to different reagents
Solution Approach 1:
The patent implements dynamics by making the overshoot parameters variable rather than fixed. The temperature controller dynamically adjusts the overshoot temperature and duration based on the specific PCR cycle step (denaturation, annealing, extension) and the thermal stability characteristics of the nucleic acid amplification enzyme used. This allows the system to adapt to different reagents and analysis items while maintaining fast temperature response.
3Reliability
If no overshooting is performed, then the reaction mixture does not overheat, but the temperature change is delayed and the reaction mixture does not reach the target temperature
Solution Approach 1:
The patent resolves this contradiction by dynamically adjusting overshoot parameters based on PCR cycle stages. During denaturation steps where high temperature is required, overshooting to 95°C accelerates the temperature response. During extension steps where precise temperature control is critical, the overshoot is reduced or eliminated to prevent enzyme deactivation. This stage-specific parameter adjustment maintains both speed and accuracy.
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 solution prevents enzyme deactivation and air bubble formation, enhances analytical performance, and allows for shorter analysis times by optimizing temperature control based on the analysis item and reagent characteristics, enabling flexible overshoot settings for different analysis items.
Implementation Method 1
the temperature of the temperature control block is raised to be equal to or higher than the target temperature for the temperature of the reaction mixture
Implementation Method 2
the nucleic acid is cooled to approximately 60° C. for annealing and extension reaction
Implementation Method 3
a temperature sensor which monitors a temperature of the holder
Data Source
AI summary
The purpose of the invention is to provide a nucleic acid analyzer that sets and executes temperature regulation, said temperature regulation matching the characteristics of analyzing items and the configuration of the analyzer, by a simple operation while preventing deterioration in analytical performance caused by partial overheating of a liquid reaction mixture to thereby improve temperature change speed and shorten analysis time. To achieve the above purpose, provided is a method wherein, in the case of performing overshooting: as a first processing, the temperature is continuously elevated until reaching a target overshoot temperature; as a second processing, after reaching the aforesaid temperature, the overshoot target temperature is maintained for a preset period of time; and, as a third processing, the temperature is continuously lowered until reaching a target temperature of the liquid reaction mixture. By executing the first to third processings, regulation is conducted so that temperature measurement values are in a trapezoidal form.