Sequencing Method Using Single Photodetector for Nucleotide Identification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nucleic acid sequencing methods using luminescence-based instrumentation are large and expensive, requiring improvements in throughput capacity and cost efficiency, particularly in detecting nucleotides with existing multi-channel detection systems.
Innovation Solution
A method utilizing a single detection filter to differentiate nucleotides by employing emitters with distinct excitation properties, where each nucleotide emits at specific wavelengths with varying peak intensities when irradiated with different excitation wavelengths, allowing for the identification of nucleotides using a single photodetector and potentially fewer detection channels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multi-channel detection systems are used to detect different nucleotides, then nucleotide identification accuracy is improved, but device complexity and cost increase
Solution Approach 1:
The patent applies universality by using a single photodetector to perform multiple detection functions. Instead of requiring separate detection channels for different nucleotides, the system uses one photodetector that detects emission from multiple emitter types (different fluorophores) by utilizing their distinct excitation and emission wavelength characteristics, thereby reducing device complexity while maintaining detection capability
Solution Approach 2:
The patent changes the detection parameters by shifting from detecting multiple wavelengths simultaneously with multiple photodetectors to detecting sequential emission at different wavelengths with a single photodetector. The system varies the excitation wavelength parameter to selectively excite different emitters and measures emission intensity at specific wavelengths, thereby simplifying the detection system while preserving nucleotide identification accuracy
2Measurement precision
If luminescence-based sequencing instrumentation is used, then nucleotide detection capability is improved, but instrument size and cost increase
Solution Approach 1:
The patent extracts the essential detection function from complex luminescence instrumentation by using a single photodetector with wavelength-selective filtering rather than full multi-channel spectral detection systems. This extraction approach retains the core capability to detect nucleotide incorporation through emitter fluorescence while removing unnecessary complexity, reducing instrument size and cost
Solution Approach 2:
The patent employs simpler, more cost-effective detection components instead of expensive, complex luminescence instrumentation. By using a single photodetector with appropriate optical filters rather than multiple specialized detectors and complex optical paths, the system achieves comparable detection capability with reduced instrument size and lower cost
3Measurement precision
If multiple detection channels are used to differentiate nucleotides, then sequencing accuracy is improved, but throughput capacity decreases
Solution Approach 1:
The patent implements periodic action by sequentially exciting different emitter types at different time points within each sequencing cycle. Instead of simultaneous multi-wavelength detection, the system alternates between exciting different fluorophores and measuring their emission, thereby maintaining accurate nucleotide identification while enabling faster data acquisition and higher throughput
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 speed and throughput capacity while reducing the complexity and cost of sequencing instrumentation by enabling the identification of nucleotides with fewer detection channels, thereby improving the efficiency and economic viability of nucleic acid sequencing.
Implementation Method 1
each nucleotide residue is identified as it is incorporated into the growing nucleic acid strand. The incorporated nucleotide is read using an appropriate label attached thereto... Detection of the label can be carried out using various methods, including luminescence spectroscopy or by other optical means. Generally, the preferred label is a fluorophore, which, after absorption of energy, emits radiation at a defined wavelength.
Implementation Method 2
The first emitter has different luminescence properties to the second emitter
Data Source
AI summary
A method for determining the sequence of a polynucleotide is described. The method comprises detecting in a sequencing reaction the incorporation of a first, second, third, or fourth nucleotide using first and second excitation wavelengths and a photodetector having a detection window comprising a range of wavelengths.