Polymerase-Linked Nucleotides for Low-Cost Precise Sequencing
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Solution Overview
Problem
Existing sequencing technologies are prohibitively expensive for widespread deployment across large populations, hindering genetic correlation studies and individualized healthcare applications.
Innovation Solution
Development of polymerase-linked nucleotides covalently attached to a catalytically active polymerase enzyme via a flexible linker, allowing non-covalent binding at the active site and incorporation into the 3′ end of a polynucleotide, with a detectable label, enabling high-resolution nucleotide sequencing through dwell time monitoring.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional sequencing platforms are used, then accurate nucleotide sequencing is achieved, but the cost is prohibitively expensive for widespread deployment
Solution Approach 1:
The patent combines the polymerase enzyme, nucleotide, and detectable label into a single conjugate unit. This merging eliminates the need for separate reagent additions and reduces the number of chemical components required, directly addressing the high cost issue while maintaining sequencing accuracy through the integrated design.
Solution Approach 2:
The polymerase-linked nucleotide serves multiple functions simultaneously: it acts as the polymerase enzyme for nucleotide incorporation, provides the nucleotide substrate itself, and includes a detectable label for signal detection. This multi-functionality reduces the number of separate reagents needed, lowering costs while maintaining measurement precision.
2Productivity
If conventional sequencing reagents are used, then nucleotide incorporation is achieved, but reagent consumption and cost are high
Solution Approach 1:
By combining the polymerase, nucleotide, and label into one conjugate, the patent reduces reagent consumption. Each conjugate molecule performs the complete function of nucleotide incorporation with its associated detection capability, eliminating the need for excess separate reagents and reducing overall substance quantity required.
Solution Approach 2:
The polymerase-linked nucleotide is self-sufficient, carrying its own detectable label and catalytic activity. Each molecule can independently perform nucleotide incorporation and signal generation without requiring additional reagents, reducing reagent consumption while maintaining high productivity.
3Measurement precision
If traditional sequencing methods are used, then sequence data is obtained, but error rates and background noise reduce accuracy
Solution Approach 1:
The patent extracts the detectable label from the bulk reagent pool and attaches it directly to each individual polymerase-nucleotide conjugate. This extraction of the detection function to the single-molecule level eliminates background noise from unincorporated labeled reagents, reducing errors and improving sequence determination accuracy.
Solution Approach 2:
The detectable label on each polymerase-linked nucleotide provides immediate feedback upon incorporation into the growing polynucleotide chain. This real-time detection at the single-molecule level allows for accurate signal registration and reduces errors by directly observing each incorporation event rather than relying on bulk measurements.
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
Enables cost-effective, high-throughput sequencing with reduced errors and reagent use, allowing for accurate nucleotide sequence determination and large-scale genetic analysis.
Implementation Method 1
a nucleotide covalently attached to a catalytically active polymerase enzyme by a flexible linker
Implementation Method 2
catalytically active polymerase enzyme
Implementation Method 3
the linker attachment to the nucleotide allows the nucleotide to bind non-covalently at the active site of the polymerase
Data Source
AI summary
Presented herein are polymerase-linked nucleotides for improved distinguishing nucleotide sequences for different nucleic acid molecules. Also presented are methods and systems using the polymerase-linked nucleotides for improved distinguishing nucleotide sequences for different nucleic acid molecules.