Self-luminescent nucleotide sequencing reduces equipment complexity
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Solution Overview
Problem
Current DNA sequencing methods, such as sequencing by synthesis, require complex equipment and multiple laser sources due to the use of cleavable fluorescent nucleotide reversible terminators or 2-channel chemistry, which increases costs and reduces efficiency.
Innovation Solution
A single-channel sequencing method utilizing self-luminescence patterns from nucleotide derivatives with different molecular labels and linkers, where the hydroxyl group at the 3′-position is protected by a reversible group, allowing for bioluminescence or chemiluminescence-based detection without external photoexcitation, enabling the differentiation of nucleotides A, T/U, and G through specific binding and cleavage reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If cleavable fluorescent nucleotide reversible terminators or 2-channel chemistry are used, then nucleotide identification is achieved, but equipment complexity and cost increase due to multiple laser sources and cameras
Solution Approach 1:
The patent extracts and removes the fluorescent labeling system and external photoexcitation requirements from the sequencing method. Instead of using fluorescent nucleotides that require laser excitation and complex detection systems, the invention uses nucleotides with inherent self-luminescence properties, eliminating the need for lasers and cameras while maintaining nucleotide identification capability
Solution Approach 2:
The patent replaces the optical detection system (lasers and cameras) with a chemical/biological detection system based on self-luminescence. The mechanical/optical components are substituted by utilizing the intrinsic luminescent properties of the nucleotide derivatives, which can be detected without external photoexcitation
2Adaptability or versatility
If multiple laser sources and cameras are used for 2-channel chemistry, then four different nucleotides can be detected, but cost increases
Solution Approach 1:
The patent makes the self-luminescence detection system universal for detecting all four nucleotide types (A, T/U, C, G) using a single detection channel. Different nucleotides are differentiated by their unique self-luminescence patterns rather than requiring separate detection systems, making the method versatile while cost-effective
Solution Approach 2:
The patent uses simple, inexpensive nucleotide derivatives with self-luminescence properties that can be detected with basic equipment. The approach replaces expensive lasers and cameras with simpler detection methods, reducing the cost of the sequencing system while maintaining functionality
3Device complexity
If self-luminescence patterns are used without external photoexcitation, then equipment requirements are reduced, but detection sensitivity may be affected
Solution Approach 1:
The patent changes the detection parameter from external light emission (fluorescence requiring laser excitation) to self-emitted light (self-luminescence). By utilizing the inherent luminescence properties of the nucleotide derivatives and optimizing detection conditions, the system maintains sensitivity while eliminating complex equipment requirements
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 simplifies the sequencing process, reduces equipment requirements, and enhances efficiency by using self-luminescence signals to identify nucleotides, potentially lowering costs and improving sequencing throughput.
Implementation Method 1
the luminescence signal used to implement the sequencing method is derived from bioluminescence
Implementation Method 2
utilizing self-luminescence patterns from nucleotide derivatives with different molecular labels and linkers, where the hydroxyl group at the 3′-position is protected by a reversible group, allowing for bioluminescence or chemiluminescence-based detection
Data Source
AI summary
Methods and kits for sequencing a nucleic acid molecule are provided, which include utilizing four different compounds that are respectively derivatives of nucleotides A, (T/U), C and G, wherein the hydroxyl at the 3′-position of sugar of each of the four compounds is protected by a reversible protecting group. Each of the four compounds comprises a first linker, a second linker, and a terminal molecular label binding to or reactive to a receptor in a detectable group comprising a luminescence-activating molecule and the receptor, the luminescence-activating molecule capable of causing emission of fluorescence in the presence of a suitable substrate. The second linkers in the four compounds are different, and can be cleaved at different conditions. After incorporating one of the four compounds into a growing chain using the target nucleic acid molecule as a template, a series of reactions is performed to cleave the second linker and the first linker, and a series of detections are performed for fluorescent signals, thereby determining the identity of the nucleotide of the incorporated compound.