Nucleic Acid Size Distribution via Proportional Fluorescent Labeling
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Solution Overview
Problem
Current methods for measuring the size distribution of nucleic acid molecules are limited by low throughput, accuracy, and repeatability, and lack the ability to quantify single DNA molecules effectively, which is crucial for clinical applications such as cancer diagnostics.
Innovation Solution
A process involving labeling nucleic acid molecules with fluorescent dyes proportional to their base pairs, combining them with fluorescent nanoparticles, and using specific wavelengths to excite and detect fluorescence spectra for precise size measurement, allowing for high-throughput and accurate determination of DNA sizes in samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent dyes are used to label nucleic acid molecules for size measurement, then measurement sensitivity is improved, but measurement precision deteriorates due to non-proportional labeling
Solution Approach 1:
The patent changes the chemical parameters of the labeling reaction by using specific intercalating dyes (e.g., YOYO-1, SYBR Green) that bind to nucleic acids in a predictable stoichiometric ratio. By controlling the dye-to-nucleic acid ratio and reaction conditions, the labeling achieves reliable proportionality where the number of dye molecules attached is directly proportional to the number of base pairs, enabling accurate size measurements through fluorescence intensity.
2Productivity
If conventional measurement methods are used, then device complexity is reduced, but productivity deteriorates due to low throughput
Solution Approach 1:
The patent replaces mechanical separation methods (such as gel electrophoresis) with an optical detection system. By using fluorescent labeling combined with widefield fluorescence microscopy or flow cytometry, the system can measure thousands of individual nucleic acid molecules in parallel, achieving high throughput without the mechanical complexity and time constraints of traditional separation-based methods.
3Measurement precision
If single molecule measurement is implemented, then measurement precision is improved, but reliability deteriorates due to difficulty in quantification
Solution Approach 1:
The patent incorporates calibration standards with known nucleic acid sizes and fluorescence intensities into the measurement system. By comparing the fluorescence intensity of unknown single molecules against these calibrated standards, the system achieves reliable quantification. The feedback loop allows for correction of variations in labeling efficiency, detection sensitivity, and instrumental parameters, ensuring consistent and reproducible size measurements across multiple samples and experiments.
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 provides reliable, rapid, and sensitive measurements of nucleic acid molecule sizes, overcoming limitations of existing techniques by enabling precise quantification and high throughput, suitable for clinical use.
Implementation Method 1
labeling the nucleic acid molecules with a fluorescent dye comprising a plurality of fluorescent dye molecules to form labeled nucleic acid molecules... irradiating the sample at a first wavelength, the first wavelength exciting the labeled nucleic acid molecules... producing, by the labeled nucleic acid molecules, the first florescence spectrum
Data Source
AI summary
A process for measuring a size distribution of a plurality of nucleic acid molecules, the process comprising: labeling the nucleic acid molecules with a fluorescent dye comprising a plurality of fluorescent dye molecules to form labeled nucleic acid molecules, such that a number of fluorescent dyes molecules attached to each nucleic acid molecule is reliably proportional to the number of base pairs in the nucleic acid molecule, the fluorescent dye molecules having a first florescence spectrum; producing, by the labeled nucleic acid molecules, the first florescence spectrum in response to irradiating the labeled nucleic acid molecules at the first wavelength; and detecting the first florescence spectrum to measure the size distribution of the plurality of nucleic acid molecules.