Sequencing Probe Compositions for Amplification-Free Nucleotide Reading
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Solution Overview
Problem
Current nucleic acid sequencing methods require enzymatic amplification and polymerization steps, which are costly and time-consuming, necessitating a need for rapid, enzyme-free, and amplification-free sequencing solutions.
Innovation Solution
The development of sequencing probes with a target binding domain and barcode domain, utilizing a synthetic backbone with attachment positions and complementary nucleic acid molecules, enabling direct nucleotide identification without amplification or enzymes, and a method for determining nucleotide sequences through hybridization and label detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If enzymatic amplification and polymerization steps are used for nucleic acid sequencing, then sequencing accuracy and signal detection are improved, but sequencing time and cost increase
Solution Approach 1:
The invention extracts and eliminates the enzymatic amplification and polymerization steps from the sequencing process. By using direct hybridization of probes to target nucleic acids followed by detection of hybridization signals, the method removes the time-consuming enzymatic steps while maintaining sequencing capability through alternative detection mechanisms.
Solution Approach 2:
The invention introduces probe molecules as intermediaries that hybridize to target nucleic acids. These probes serve as mediators between the target sequence and the detection system, enabling signal generation through hybridization events rather than enzymatic reactions, thus achieving rapid sequencing without enzymes.
2Measurement precision
If enzymatic amplification steps are used for nucleic acid sequencing, then detectable signal is improved, but cost and time consumption increase
Solution Approach 1:
The invention extracts and eliminates the enzymatic amplification steps from the sequencing process. By using direct hybridization of probes to target nucleic acids followed by detection of hybridization signals, the method removes the time-consuming enzymatic steps while maintaining sequencing capability through alternative detection mechanisms.
Solution Approach 2:
The invention performs preliminary hybridization of multiple probes to different regions of the target nucleic acid simultaneously. This preliminary binding establishes multiple detection points in parallel, enabling high-throughput sequencing without the need for sequential enzymatic amplification steps.
3Reliability
If conventional sequencing methods with amplification steps are used, then sequencing completeness is improved, but process complexity increases
Solution Approach 1:
The invention segments the target nucleic acid into multiple regions that can be simultaneously bound by different probes. Each probe targets a specific segment, and the collective data from all probe hybridizations provides complete sequencing information. This segmentation approach simplifies the overall process by enabling parallel detection across multiple regions.
Solution Approach 2:
The invention uses universal probe structures that can hybridize to various target sequences through complementary base pairing. The probes serve multiple functions: target recognition, signal generation, and sequence determination. This multi-functionality reduces process complexity by eliminating the need for separate amplification and detection steps.
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 rapid, enzyme-free, and amplification-free nucleic acid sequencing with long read lengths and low error rates, suitable for clinical applications.
Implementation Method 1
the target binding domain hybridizes to a target nucleic acid
Implementation Method 2
each attachment position comprising at least one nucleic acid sequence that hybridizes to a complementary nucleic acid molecule
Data Source
AI summary
The present disclosure relates to chemical compositions, kits, and apparatuses and methods for using these compositions, kits and apparatuses in various assays.