Chromenoquinoline Dye Composition for Stable Multiplex Sequencing
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Solution Overview
Problem
Existing fluorescent dyes for nucleic acid sequencing face challenges in achieving spectrally-distinguishable absorption and emission spectra, compatibility with biological reagents, and stability under high pH conditions, limiting multiplex detection efficiency and throughput.
Innovation Solution
Development of chromenoquinoline dyes with long Stokes shifts and improved fluorescent intensity, excitable by blue and green light, and enhanced stability in high pH buffers, suitable for nucleotide labeling and sequencing applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional fluorescent dyes are used for multiplex detection, then spectral resolution may be achieved, but the dyes lack compatibility with aqueous reagents and enzymes, reducing reliability
Solution Approach 1:
The patent modifies the chemical structure of chromenoquinoline dyes by changing parameters such as introducing hydrophilic groups (carboxyl, sulfonate) and adjusting molecular weight and charge characteristics. These parameter changes enable the dyes to be compatible with aqueous reagents, buffers, and enzymes while maintaining their fluorescent properties and spectral resolution capabilities.
2Measurement precision
If dyes with resolved absorption and emission spectra are used, then multiplex detection is enabled, but photo-stability under high power laser excitation deteriorates
Solution Approach 1:
The patent creates composite fluorescent probes by combining chromenoquinoline core structures with stabilizing moieties and protective groups. The dyes are conjugated to nucleotides and other molecules in composite structures that enhance photo-stability while maintaining the resolved absorption and emission spectra necessary for multiplex detection under high power laser excitation.
3Measurement precision
If blue light excitation is used to improve optical resolution, then imaging resolution improves, but dye stability and fluorescence intensity under these conditions deteriorate
Solution Approach 1:
The patent specifically designs chromenoquinoline dyes with modified molecular parameters including adjusted HOMO-LUMO gaps, introduced hydrophilic substituents, and optimized molecular weights that enable stable fluorescence under blue light excitation (450-460 nm). These parameter changes allow the dyes to maintain both high optical resolution and dye stability when excited by blue light sources.
4Measurement precision
If fluorescent intensity is increased for better detection, then sequencing accuracy improves, but compatibility with sequencing reagents and enzymes deteriorates
Solution Approach 1:
The patent applies local quality modifications by introducing specific functional groups (carboxyl, sulfonate, hydroxyl) at particular positions on the chromenoquinoline molecule. These localized modifications provide reagent compatibility and enzyme compatibility in specific regions while preserving the overall high fluorescence intensity needed for accurate sequencing detection.
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
The chromenoquinoline dyes provide strong fluorescence and chemical stability, enabling efficient multiplex detection and higher throughput in nucleic acid sequencing, reducing sequencing errors and reagent usage.
Implementation Method 1
chromenoquinoline dyes with long Stokes shifts and improved fluorescent intensity and chemical stability, excitable by both blue and green light
Implementation Method 2
chromenoquinoline dyes with long Stokes shifts and improved fluorescent intensity and chemical stability
Implementation Method 3
covalently attached to photo-protecting cyclooctatetraene moieties for enhanced performance
Data Source
AI summary
The present application relates to chromenoquinoline dyes and their uses as fluorescent labels. For example, these dyes may be used to label nucleotides for nucleic acid sequencing.