Multiplex Sequencing Signal Separation for Higher Throughput
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current sequencing methods, such as sequencing by synthesis (SBS) and sequencing by binding (SBB), are limited by the interference of simultaneous sequencing signals at different sites, leading to reduced sequencing speed and data throughput.
Innovation Solution
A method that allows simultaneous sequencing of multiple sequencing templates at one position by detecting and distinguishing sequencing signals based on intensity differences, using a sequencing library with dual-indexing and nucleic acid molecules with specific barcodes, enabling synchronous sequencing of adapters at both ends.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple sequencing templates are sequenced simultaneously at one position, then sequencing speed and data throughput are improved, but signal interference and confusion occur between simultaneous sequencing signals
Solution Approach 1:
The patent introduces blocking molecules as intermediaries that selectively bind to primer hybridization sites on certain nucleic acid molecules, preventing them from being sequenced in specific cycles. This mediator allows the system to control which templates are active in each cycle, resolving the signal interference problem while maintaining high throughput by enabling multiple templates to be sequenced across different cycles at the same position
Solution Approach 2:
The patent implements periodic sequencing cycles where different sets of nucleic acid molecules are sequenced in alternating cycles. By using blocking molecules to periodically activate or deactivate specific templates, the system achieves periodic action that prevents simultaneous signal interference while maintaining overall sequencing productivity through systematic rotation of active templates
2Measurement precision
If conventional sequencing methods are used with single indexing, then sequencing signals can be clearly distinguished, but sequencing efficiency is limited and adapters at both ends cannot be sequenced simultaneously
Solution Approach 1:
The patent merges the sequencing of adapters at both ends of nucleic acid molecules into a single sequencing run by using dual-indexing with blocking molecules. This combining approach allows both adapters to be sequenced simultaneously in the same cycle without requiring separate sequencing runs, thereby improving efficiency while maintaining signal distinction through the blocking mechanism
Solution Approach 2:
The patent creates a universal sequencing system where a single sequencing run can handle multiple functions: sequencing both adapters at the ends of nucleic acid molecules, processing multiple different templates simultaneously, and maintaining signal distinction. The dual-indexing system with blocking molecules provides multi-functionality that increases productivity without sacrificing measurement precision
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 sequencing efficiency by allowing simultaneous sequencing of multiple templates, improving speed and throughput by retaining and classifying sequencing signals from different templates, thus overcoming the limitations of conventional methods.
Implementation Method 1
a first barcode, including a first primer hybridization site and a first barcode sequence ligated with a 3' end of the first primer hybridization site
Data Source
Figure 1~3
Figure 4~5
Figure 6~8
AI summary
Provided is a sequencing method. The sequencing method comprises: detecting sequencing signals of a specified position on the surface of a solid-phase support in a sequencing reaction process, the specified position comprising at least two sequencing signals; and on the basis of the difference between the sequencing signals, acquiring the type of the base at the specified position, thereby achieving the sequencing of the specified position. The method achieves the sequencing of multiple signal sites, thus increasing the sequencing speed of the sequencing reaction and the sequencing throughput per unit time.