Self-Luminescent Single-Channel Sequencing Without External Optics
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Solution Overview
Problem
Current DNA sequencing technologies, particularly single-channel methods, are costly due to the need for external light sources and additional designs to filter background light, and suffer from high error rates and instrument bulkiness, limiting their miniaturization and portability.
Innovation Solution
A self-luminescence-based sequencing method using luciferase-ligated nucleotides, eliminating the need for external light sources and additional filtering, and ensuring accurate sequencing by blocking the 3'-hydroxyl group to allow only one nucleotide incorporation per reaction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If external light sources and filtering designs are used to detect fluorescent signals, then sequencing accuracy is improved, but device complexity and cost increase
Solution Approach 1:
The patent extracts and removes the external light source and filtering components from the sequencing device. By using fluorescent dyes that inherently emit light without external excitation, the complex optical system (lasers, filters, cameras) is eliminated, retaining only the essential detection function while dramatically simplifying device structure.
Solution Approach 2:
The fluorescent dyes used in the patent possess self-luminescence capability, meaning they can emit light signals autonomously without requiring external light sources for excitation. This self-service characteristic eliminates the need for complex optical excitation systems while maintaining signal detection accuracy.
2Difficulty of detecting and measuring
If external light sources are used for fluorescence detection, then signal detection capability is improved, but instrument volume increases
Solution Approach 1:
The patent removes the bulky external light source components (lasers, optical paths, filters) from the instrument. By relying on the self-luminescent property of fluorescent dyes, the detection system can be miniaturized to essential components only, dramatically reducing instrument volume while preserving signal detection capability.
Solution Approach 2:
The fluorescent dyes generate their own light signals without external excitation, eliminating the need for large optical systems. This self-service mechanism enables the detection function to be achieved with minimal components, facilitating instrument miniaturization and portability.
3Measurement precision
If multiple fluorescent dyes are used to differentiate 4 bases, then base identification accuracy is improved, but manufacturing cost increases
Solution Approach 1:
The patent applies different fluorescent dyes with distinct emission characteristics to different nucleotide bases (A, T, C, G). Each base is labeled with a specific fluorescent dye that emits at a characteristic wavelength, enabling base differentiation through spectral analysis. This local quality differentiation achieves accurate base identification while using cost-effective fluorescent labeling strategies.
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 approach reduces sequencing costs, enhances accuracy, and enables miniaturization and portability of sequencing devices by utilizing bioluminescence or chemiluminescence signals without external excitation, facilitating on-site detection.
Implementation Method 1
adding two different luciferases to carry out a binding reaction... adding a substrate of the first luciferase and detecting luminescence signal at the same time... adding a substrate of the second luciferase and detecting luminescence signal at the same time
Implementation Method 2
it is usually necessary to use 4 kinds of fluorescent dyes to respectively label these 4 bases... in order to read the fluorescent signal carried by each base
Data Source
Figure 1
AI summary
The present invention provides a sequencing method based on a single fluorescent dye, in which a self-luminescence signal is used to distinguish the sequential incorporation of different nucleotides, thereby realizing the determination of the polynucleotide sequence.