Modified Nucleotide Linkers for Sequencing by Synthesis
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Solution Overview
Problem
Current nucleotide incorporation methods in DNA sequencing by synthesis are limited by slow nucleotide incorporation rates, which hinder the efficiency of sequencing processes.
Innovation Solution
Development of nucleosides or nucleotides covalently attached to fluorophores through specific linkers, such as structures represented by formulas (I) and (II), which enhance nucleotide incorporation rates by modifying the linker structure to improve sequencing efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional nucleotide linkers are used in sequencing by synthesis, then the sequencing process can proceed, but the nucleotide incorporation rate is slow, limiting sequencing efficiency
Solution Approach 1:
The patent modifies the chemical structure of the linker by changing parameters such as the introduction of sulfur atoms (forming disulfide bonds), adjustment of chain length (m, n, p values), and substitution patterns (R1-R8 groups) to optimize the nucleotide incorporation rate while maintaining sequencing functionality
Solution Approach 2:
The linker is designed as a composite structure combining multiple functional elements: fluorophore attachment group, nucleotide attachment group, and intermediate chain with specific substituents (R1-R8), creating a multifunctional molecule that simultaneously enables detection and efficient incorporation
2Reliability
If standard linkers are used, then the sequencing process is simple, but phasing and pre-phasing errors occur, reducing sequencing accuracy
Solution Approach 1:
The patent introduces specific functional groups at localized positions within the linker (R1-R8 substituents at different locations) to perform specific functions such as reducing phasing errors, maintaining structural stability, and enabling accurate base identification without complicating the overall sequencing process
Solution Approach 2:
The modified linker acts as an intermediary between the fluorophore and the nucleotide, mediating their interaction to prevent phasing and pre-phasing errors while maintaining the essential sequencing function, thereby improving accuracy without requiring fundamental changes to the sequencing system
Data Source
AI summary
Some embodiments of the present application relate to novel modified nucleotide linkers for increasing the efficiency of nucleotide incorporation in Sequencing by Synthesis applications. Methods of preparing these modified nucleotide linkers are also provided herewith.