Single-Nucleotide Multi-Fluorophore Complexes for Signal-Enhanced Sequencing
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Solution Overview
Problem
Current nucleic acid sequencing methods, particularly sequencing-by-synthesis (SBS), are limited by the use of single fluorophores per nucleotide, which restricts sensitivity and signal intensity.
Innovation Solution
The incorporation of nucleotide-fluorophore complexes, where each complex comprises a single nucleotide attached to a core with a plurality of fluorophores, enhances signal intensity and sensitivity by allowing multiple fluorophores to be associated with each nucleotide.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If a single fluorophore is used per nucleotide in sequencing-by-synthesis, then the method is simple and easy to implement, but the signal intensity and sensitivity are limited
Solution Approach 1:
The patent combines multiple fluorophores with a single nucleotide to form a nucleotide-fluorophore complex. This merging of multiple fluorophores onto one nucleotide structure increases the signal intensity while maintaining the sequencing-by-synthesis process, directly resolving the contradiction between signal intensity and method simplicity.
Solution Approach 2:
The invention creates a composite nucleotide-fluorophore complex where multiple fluorophores are integrated with a single nucleotide. This composite structure allows the system to achieve higher signal intensity without fundamentally changing the sequencing methodology, thus addressing the contradiction between improved signal and maintained simplicity.
2Measurement precision
If multiple fluorophores are associated with each nucleotide, then signal intensity and sensitivity are improved, but the complexity of the nucleotide structure increases
Solution Approach 1:
By merging multiple fluorophores with a single nucleotide into a unified complex structure, the patent achieves improved sensitivity without requiring separate handling of multiple nucleotides. The merged complex is incorporated as a single unit during sequencing, maintaining process simplicity while enhancing measurement precision.
3Measurement precision
If multiple fluorophores are attached to each nucleotide, then signal-to-noise ratio improves, but the manufacturing complexity increases
Solution Approach 1:
The patent employs preliminary action by pre-assembling the nucleotide-fluorophore complexes before the sequencing process. The complexes are prepared in advance with multiple fluorophores attached to each nucleotide, allowing for optimized signal-to-noise ratio while simplifying the actual sequencing manufacturing process, as the complexes are ready for direct incorporation.
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 improves signal intensity and signal-to-noise ratios, leading to increased sensitivity in nucleic acid sequencing.
Implementation Method 1
each nucleotide is labelled with a single fluorophore which can limit sensitivity and signal intensity
Data Source
AI summary
The present disclosure relates in some aspects to methods, systems, and kits for sequencing a template nucleic acid molecule using nucleotide-fluorophore complexes. In some embodiments, each nucleotide-fluorophore complex comprises: (i) a single nucleotide attached to a core; and (ii) a plurality of fluorophores attached to the core; wherein each nucleotide-fluorophore complex comprises no more than one nucleotide. In some embodiments, the methods provided herein achieve improved signal intensity and sensitivity.