Cyclic Reducing Agents for SBS Sequencing Accuracy

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Solution Overview

Problem

Sequencing by synthesis (SBS) technologies face challenges due to residual reducing agents that prematurely cleave reversible terminators and labels, leading to asynchronous nucleotide incorporation and lower sequencing accuracy.

Innovation Solution

The use of cyclic reducing agents with a unique polymeric structure that can be easily removed from the reaction vessel, allowing for controlled nucleotide incorporation and improved sequencing accuracy by preventing premature cleavage of reversible terminators and labels.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional reducing agents are used in SBS sequencing, then nucleotide incorporation can proceed, but residual reducing agents prematurely cleave reversible terminators and labels leading to sequencing errors

Engineering Contradiction:

Improvesequencing cycle timeVSAvoidsequencing accuracy

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent removes the harmful residual reducing agent from the reaction system by using a solid-supported reducing agent that can be easily separated from the solution after the reduction step, preventing premature cleavage of reversible terminators and labels in subsequent sequencing cycles

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces a solid-supported reducing agent as an intermediary that performs the necessary reduction function while being physically separable from the reaction mixture, thus mediating between the need for reduction and the need to avoid residual interference

Inventive Principle:

Principle #24Intermediary (Mediator)

2Duration of action of moving object

If reducing agents remain in the reaction vessel, then nucleotide incorporation continues, but reversible terminators and labels are prematurely cleaved causing dephasing

Engineering Contradiction:

Improvenucleotide incorporation durationVSAvoidsynchronization of nucleotide incorporation

Core Design Contradiction:

Duration of action of moving objectVSManufacturing precision

Solution Approach 1:

The solid-supported reducing agent is extracted from the reaction mixture after performing its reduction function, eliminating the source of premature cleavage and maintaining synchronization across the nucleotide incorporation population

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent employs a disposable solid-supported reducing agent that is used for a single reduction cycle and then discarded, ensuring no residual activity carries over to subsequent sequencing cycles where it would cause dephasing

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

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 results in faster sequencing cycle times, reduced out-of-phase values, and longer sequencing read lengths by maintaining the integrity of nucleotide incorporation and detection processes.

Implementation Method 1

contacting the incorporated nucleotide analogues with a compound as described herein, including embodiments, to remove the detectable label

Methodology Applied

Scientific EffectReduction: Reduction

Implementation Method 2

contacting the nucleotide analogue with a compound as described herein, including embodiments, to remove the reversible terminator

Methodology Applied

Scientific EffectReduction: Reduction

Data Source

PatentUS20240368202A1Reducing agents and uses thereof

Publication Date: 2024.11.07 SINGULAR GENOMICS SYSTEMS INC

AI summary

Disclosed herein, inter alia, are reducing agents and methods of use thereof.