Electrode-Based DNA Bridge Detection via Rolling Circle Amplification
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Solution Overview
Problem
Conventional DNA detection methods require amplification and struggle with detecting short DNA sequences due to limitations in nano-tweezers miniaturization, and they often rely on marker substances like fluorescent reagents.
Innovation Solution
A method using a detection device with electrodes to immobilize primers, generate DNA bridges through Rolling Circle Amplification, and characterize them using resonance frequency, electrical conductance, or nanoparticle coating without the need for labeling substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Difficulty of detecting and measuring
If conventional nano-tweezers are miniaturized to detect short DNA, then detection capability for short DNA improves, but manufacturing precision and structural stability deteriorate due to miniaturization limits
Solution Approach 1:
The patent replaces the mechanical nano-tweezers system with an electrode-based system that uses electrical fields and Rolling Circle Amplification to detect DNA. This substitution avoids the miniaturization limits of mechanical structures while maintaining high detection capability for short DNA sequences.
Solution Approach 2:
The patent performs preliminary amplification of DNA using Rolling Circle Amplification before detection. This preliminary action increases the DNA signal strength, enabling detection of short DNA sequences without requiring extreme miniaturization of the detection device itself.
2Measurement precision
If DNA amplification is performed before detection, then detection sensitivity improves, but detection time and process complexity increase
Solution Approach 1:
The patent performs preliminary amplification of DNA using Rolling Circle Amplification before detection. This preliminary action increases the DNA signal strength, enabling detection of short DNA sequences without requiring extreme miniaturization of the detection device itself.
Solution Approach 2:
The patent changes the physical parameters of DNA detection by measuring electrical conductance and resonance frequency of DNA bridges between electrodes. These parameter changes enable direct detection of amplified DNA products, reducing the time needed for additional processing steps.
3Reliability
If marker substances like fluorescent reagents are used, then detection reliability improves, but device complexity and cost increase
Solution Approach 1:
The patent enables DNA to serve its own detection function by measuring its intrinsic electrical conductance and resonance frequency properties. This self-service approach eliminates the need for external markers or labels, simplifying the device while maintaining reliable detection.
Solution Approach 2:
The patent replaces optical detection methods requiring fluorescent markers with electrical detection methods that measure the conductance and resonance frequency of DNA bridges. This substitution eliminates marker substances while maintaining detection reliability.
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
Enables efficient and marker-free detection of DNA by forming and characterizing DNA bridges between electrodes, overcoming the limitations of miniaturization and amplification requirements, and allowing real-time detection.
Implementation Method 1
generating single stranded DNA product utilizing RCA (Rolling Circle Amplification), with impressing a designated voltage between the electrodes
Implementation Method 2
characterizing the bridge of DNA is based on a resonance frequency of the bridge of DNA between the electrodes
Implementation Method 3
characterizing the bridge of DNA is based on an electrical conductance of the bridge of DNA between the electrodes
Implementation Method 4
coating the bridge of the DNA with conductive nanoparticles; and verifying existence of the bridge of DNA between the electrodes
Data Source
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AI summary
An object is to make a bridge of DNA expanding between a pair of electrodes, and to characterize the bridge of DNA, to thereby detect DNA easily and surely without employing any marker or labeling substances, such as fluorescent reagents. A method of detecting DNA using a detection device with at least a couple of electrodes, the method comprising immobilizing a primer on the electrodes; making a bridge of the DNA expanded between the electrodes, by immersing the electrodes in a solution including circular templates of single stranded DNA, annealing the circular templates, and generating single stranded DNA product utilizing RCA (Rolling Circle Amplification), with impressing a designated voltage between the electrodes; and characterizing the bridge of DNA which includes multiple single stranded DNA molecules between the electrodes.