Nucleic Acid Sequencing Consensus Method
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current single molecule sequencing technologies face challenges in achieving high accuracy, particularly in medical diagnostics and healthcare, due to low raw read accuracy and complex manipulation procedures that limit their applicability.
Innovation Solution
A method involving hybridization of capture primers with target nucleic acid molecules, nascent strand synthesis, and single-molecule sequencing, followed by bioinformatics analysis to generate consensus sequences, which simplifies detection steps, reduces time, and improves accuracy by immobilizing template molecules on a sequencing chip and using a predetermined similarity threshold.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If circular template method is used to improve sequencing accuracy, then sequencing accuracy is enhanced to meet application requirements, but manipulation procedures become complex and operation time becomes lengthy
Solution Approach 1:
The patent segments the sequencing process into distinct stages: template preparation, sequencing reaction, and data analysis. By using linear templates with specific adapter sequences rather than requiring circularization, the method simplifies the template preparation step while maintaining the ability to generate high-accuracy consensus sequences through multiple reads of the same template
Solution Approach 2:
The patent performs preliminary actions by pre-preparing templates with integrated adapter sequences and barcodes during the library preparation phase. This eliminates the need for subsequent circularization steps and enables direct sequencing, thereby reducing manipulation complexity while preserving accuracy through consensus calling
2Measurement precision
If circular template method is used to improve sequencing accuracy, then sequencing accuracy is enhanced to meet application requirements, but operation time becomes lengthy
Solution Approach 1:
The patent performs preliminary actions by pre-preparing templates with integrated adapter sequences and barcodes during the library preparation phase. This eliminates the need for subsequent circularization steps and enables direct sequencing, thereby reducing manipulation complexity while preserving accuracy through consensus calling
Solution Approach 2:
The patent skips the time-consuming circularization step by using linear templates with adapter sequences that enable direct sequencing. The method rushes through the process by performing sequencing directly on linear templates, generating sufficient read depth for accurate consensus sequences without waiting for circular template formation
3Speed
If single molecule sequencing is used to achieve fast sequencing speed, then sequencing speed reaches up to 5 bases per second, but raw read accuracy remains barely at 85%
Solution Approach 1:
The patent applies feedback by implementing quality control metrics that monitor raw read accuracy in real-time. Templates that fail to meet minimum quality thresholds are identified and excluded from consensus sequence generation, ensuring that only high-quality reads contribute to the final accurate sequence while maintaining fast sequencing speeds
Solution Approach 2:
The patent uses copying by generating multiple independent reads of the same template molecule and creating consensus sequences through alignment and comparison. This redundancy allows the system to overcome individual read errors while maintaining the speed advantage of single-molecule sequencing, achieving both high speed and high accuracy
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 enhances detection accuracy and reduces time, enabling broader application of single molecule sequencing by improving the sequencing process through simplified steps and bioinformatics analysis.
Implementation Method 1
hybridizing first capture primers to the first target nucleic acid molecules to form first capture primer-first target nucleic acid molecule complexes
Implementation Method 2
performing a nascent strand synthesis reaction to form first template molecule-first target nucleic acid molecule complexes
Implementation Method 3
sequencing the first template molecules by single-molecule sequencing to obtain first original sequences
Data Source
AI summary
A method for generating high accuracy sequencing results from low accuracy continuous single molecule sequencing includes the following steps: providing a sample including first target nucleic acid molecules; hybridizing first capture primers to the first target nucleic acid molecules to form first capture primer-first target nucleic acid molecule complexes; performing a nascent strand synthesis reaction to form first template molecule-first target nucleic acid molecule complexes; dissociating first template molecules; hybridizing first sequencing primers to the dissociated first template molecules, and sequencing the first template molecules by single-molecule sequencing to obtain first original sequences; comparing the first original sequences and a first target sequence to calculate a first sequence similarity of each of the first original sequences; collecting first original sequences whose first sequence similarity reaches a predetermined threshold to obtain a first cluster; and analyzing the first cluster by bioinformatics to obtain a first consensus sequence.