Fluorescent Probe Hybridization for Faster Nucleic Acid Index Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Second-generation sequencing technologies face high costs and long times due to the need to sequence indexes using SBS, which is inefficient and time-consuming.
Innovation Solution
A method for nucleic acid sequence detection involving hybridization with fluorescently labeled probes to directly identify index sequences, eliminating the need for index sequencing, using a method that includes immobilizing nucleic acids with preset sequences on a solid support, hybridizing probes with fluorescent labels, and collecting fluorescence signals to identify sequence information.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If SBS sequencing is used to sequence indexes, then sequence information can be obtained, but sequencing time and cost increase significantly
Solution Approach 1:
The patent uses fluorescently labeled probes as copies of the index sequences. Instead of sequencing the index directly, the probe serves as a pre-synthesized complementary copy that can be directly detected through fluorescence, eliminating the need for time-consuming SBS sequencing of the index
Solution Approach 2:
The patent replaces the mechanical SBS sequencing process with a biochemical hybridization system. The fluorescent probe binds to the index sequence through complementary base pairing, and the fluorescence signal provides direct detection without requiring the complex mechanical synthesis and detection cycles of SBS
2Measurement precision
If SBS sequencing is used to sequence indexes, then sequence information can be obtained, but sequencing cost increases
Solution Approach 1:
The patent uses fluorescently labeled probes as copies of the index sequences. Instead of sequencing the index directly, the probe serves as a pre-synthesized complementary copy that can be directly detected through fluorescence, eliminating the need for time-consuming SBS sequencing of the index
Solution Approach 2:
The patent employs inexpensive fluorescent probes that can be easily synthesized and discarded after use. These probes are much cheaper than the SBS sequencing reagents, and their short operational lifetime (single-use hybridization) makes them economically viable for routine indexing
3Reliability
If index sequencing is performed, then sample identification is achieved, but overall sequencing efficiency decreases
Solution Approach 1:
The patent performs preliminary action by pre-synthesizing the fluorescent probes before the actual sequencing experiment. The probes are prepared in advance with known sequences and fluorescent labels, so that during the experiment, only simple hybridization and fluorescence detection are needed, rather than performing complex sequencing for each sample
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
Significantly shortens sequencing time and reduces costs by allowing direct identification of index sequences through fluorescence, eliminating the need for index sequencing, thus improving efficiency and reducing overall sequencing time and expenses.
Implementation Method 1
detecting sequence information of the nucleic acid through a hybridization signal between a probe and the nucleic acid
Implementation Method 2
the probe that is reversely complementary to the M type(s) of the nucleic acid with the preset sequence each carries a preset fluorescent label
Data Source
AI summary
The present disclosure discloses a method for nucleic acid sequence detection and a sequencing method. The method for nucleic acid sequence detection includes: providing a nucleic acid with a preset sequence; and detecting sequence information of the nucleic acid through a hybridization signal between a probe and the nucleic acid.