Fluorescent-Label Sequencing for High-Accuracy Sequence Readout
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
High-throughput sequencers suffer from sequencing errors due to chemical, optical, or software noise, which cannot be identified at a single readout site and are not effectively eliminated, limiting the accuracy of sequence information.
Innovation Solution
A method involving the use of sequencing reagents with different nucleotide monomers conjugated to labels that become fluorescent after incorporation into the target polynucleotide, allowing for sequence information to be obtained through fluorescence detection, and multiple rounds of sequencing with varying reagent combinations to reduce or eliminate errors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high-throughput sequencing is performed using conventional SBS methods with single-base readout, then large amounts of sequence information can be obtained, but sequencing errors accumulate and accuracy decreases
Solution Approach 1:
The sequencing process is divided into multiple discrete cycles, each incorporating only 1-2 specific nucleotide types with fluorescent labels. By segmenting the four nucleotide types into separate incorporation rounds (e.g., A/T in first round, C/G in second round), the method reduces error accumulation while maintaining high-throughput capability through systematic multi-cycle sequencing
Solution Approach 2:
Nucleotide monomers are pre-conjugated with fluorescent labels before sequencing begins. This preliminary labeling allows for direct detection of incorporated nucleotides without post-incorporation modification, reducing signal acquisition errors and enabling real-time monitoring of each incorporation event across multiple sequencing cycles
2Measurement precision
If multiple rounds of sequencing are performed to eliminate errors through deep sequencing, then accuracy improves, but the complexity of the sequencing process increases
Solution Approach 1:
The sequencing system uses universal fluorescent labels that can be detected across all sequencing cycles using the same detection methodology. The same polynucleotide replicating catalyst and label detection system are reused in each cycle, allowing multiple rounds of sequencing to be performed with consistent parameters, thereby improving accuracy without proportionally increasing system complexity
Solution Approach 2:
The method systematically changes the parameter of nucleotide type incorporation across cycles (e.g., cycling through different combinations of A, T, C, G) while keeping other parameters constant. This controlled parameter variation enables error elimination through multiple reads without requiring fundamental changes to the sequencing apparatus or protocol structure
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
Enhances sequencing accuracy by reducing or eliminating errors, achieving a code accuracy rate of at least 99% and enabling read lengths up to 2400 base pairs.
Implementation Method 1
the nucleotide monomer/first label conjugates are substantially non-fluorescent until after incorporation of the nucleotide monomer into the target polynucleotide based on complementarity to the target polynucleotide
Data Source
Figure 1
Figure 2
Figure 3~4
AI summary
A method for sequencing a biological molecule, such as a nucleic acid molecule, and a method for detecting and/or correcting sequencing error(s) in the sequencing results are provided. Kits and systems based on the above methods are also provided.