Nucleic Acid Detection Dye with Peptide Modifier
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Solution Overview
Problem
Current fluorescent nucleic acid-binding dyes have limited sensitivity due to high background fluorescence, which restricts the detection of small amounts of nucleic acids and can inhibit PCR reactions, especially in real-time PCR applications.
Innovation Solution
A reagent composition comprising a light-emitting dye capable of binding to nucleic acids and a dye modifier that interacts with the dye to reduce background fluorescence and enhance sensitivity, allowing for linear detection over a wider range without inhibiting nucleic acid amplification reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent dyes are used to detect nucleic acids, then detection capability is provided, but background fluorescence increases and sensitivity decreases
Solution Approach 1:
A cationic peptide is introduced as an intermediary substance that binds to the fluorescent dye and suppresses its background fluorescence. The peptide acts as a mediator between the dye and the detection system, reducing harmful background signals while preserving the dye's ability to bind nucleic acids and provide detection capability.
Solution Approach 2:
The invention changes the physical and chemical parameters of the fluorescent dye by combining it with a cationic peptide. This combination alters the fluorescence properties of the dye, specifically reducing background fluorescence intensity while maintaining nucleic acid binding affinity, thereby improving detection sensitivity.
2Measurement precision
If dye concentration is increased to improve signal strength, then detection sensitivity improves, but PCR inhibition increases
Solution Approach 1:
The cationic peptide serves as a carrier that delivers the fluorescent dye into the PCR reaction mixture. This intermediary system allows the dye to function at optimized concentrations that provide sufficient signal strength without causing excessive PCR inhibition, as the peptide facilitates controlled dye distribution and binding.
Solution Approach 2:
By combining the dye with a cationic peptide, the invention changes the effective concentration and distribution parameters of the dye in the reaction mixture. This allows maintaining adequate signal intensity while reducing the inhibitory effect on PCR, as the peptide-modified dye exhibits different kinetic and interaction properties than free dye.
3Measurement precision
If traditional nucleic acid dyes are used, then binding affinity is provided, but sensitivity is limited and background fluorescence is high
Solution Approach 1:
The invention creates a composite system combining a fluorescent dye with a cationic peptide. This composite material exhibits enhanced properties compared to the individual components: the peptide-dye conjugate maintains nucleic acid binding affinity while suppressing background fluorescence, achieving improved detection sensitivity without the harmful background signals of traditional dyes.
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
The composition provides significantly improved sensitivity and a wider linear detection range for nucleic acids, enabling detection from as low as 1 ng/mL to 1000 ng/mL and minimizing PCR inhibition, thus enhancing the accuracy and reliability of nucleic acid quantitation and amplification processes.
Implementation Method 1
Fluorescent dyes or stains can be used in the detection of nucleic acids... These dyes can be used to detect and quantify DNA and RNA... the dye is capable of binding to a nucleic acid... the first complex is optically detectable at a wavelength in a range from 350 nm to 1200 nm
Implementation Method 2
the binding affinity of the dyes is enhanced due to increased electrostatic interaction between the dyes and nucleic acids
Implementation Method 3
suppressing the background or intrinsic fluorescence of a nucleic acid binding dye... a light-emitting dye modifier that interacts with said dye... the reagent composition produces a detectable signal that is linearly proportional to the amount of nucleic acids
Data Source
AI summary
The present invention relates to dyes in general. The present invention provides a wide range of dyes and kits containing the same, which are applicable for labeling a variety of biomolecules such as nucleic acids, cells and microorganisms. The present invention also provides various methods of using the dyes for research and development, forensic identification, environmental studies, diagnosis, prognosis, and/or treatment of disease conditions.