UV-Immobilized Nucleic Acid Probes Without Substrate Pretreatment
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Solution Overview
Problem
Existing methods for immobilizing nucleic acid probes on solid substrates often require pretreatment, such as amine or thiol functionalization, and can cause damage to the probes, leading to reduced immobilization efficiency and increased costs.
Innovation Solution
A method involving a nucleic acid probe with a terminus anchor chain portion composed of specific nucleotide sequences, such as stretches of Thymine or Uracil with intermediate Cytosine, immobilized using UV light without prior substrate treatment, enhancing bonding efficiency and reducing probe detachment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional UV crosslinking methods are used to immobilize nucleic acid probes, then immobilization efficiency can be improved, but probe damage increases significantly
Solution Approach 1:
The patent changes the wavelength parameter of UV light from conventional short wavelengths (254 nm) to longer wavelengths (300-500 nm, preferably 365 nm). This parameter change reduces the energy per photon, thereby reducing probe damage while maintaining immobilization efficiency through the photoreactivity of the homo-oligomer tail at these wavelengths
Solution Approach 2:
The patent introduces a homo-oligomer tail (intermediary component) composed of 7-100 nucleotides of a single base type at the terminus of the probe. This tail acts as a mediator that absorbs UV energy and facilitates crosslinking to the substrate, enabling effective immobilization while protecting the main probe sequence from direct UV exposure and damage
2Reliability
If amine or thiol functionalization is performed on substrates before probe immobilization, then probe attachment can be improved, but process complexity and cost increase
Solution Approach 1:
The patent extracts and eliminates the substrate pretreatment step (amine or thiol functionalization) from the immobilization process. By using UV-crosslinkable homo-oligomer tails that can directly interact with unmodified substrates, the method removes the complex multi-step functionalization process while maintaining effective probe attachment
Solution Approach 2:
The nucleic acid probe with homo-oligomer tail is self-sufficient for immobilization. The tail contains all necessary photoreactive groups to crosslink directly to the substrate upon UV exposure, eliminating the need for the substrate to provide functional groups or undergo pretreatment
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 method achieves high immobilization efficiency with low risk of probe damage, allowing cost-effective production of nucleic acid probes on thermoplastic substrates without pretreatment, suitable for applications like PCR and microfluidic devices.
Implementation Method 1
anchoring the anchor chain portion of the nucleic acid probe to the solid support by subjecting it to UV light
Data Source
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AI summary
Disclosed are nucleic acid probes, a method of immobilizing the nucleic acid to a solid support, a solid support comprising an immobilized nucleic acid probes, and a test device comprising a solid support. The nucleic acid probe includes a terminus anchor chain portion, and a capture portion wherein the terminus anchor chain portion includes a sequence of at least 18 nucleotides composed of stretches of up to 5 nucleotides of base type X with intermediate nucleotide(s) of base type Cytosine (C) and optionally one nucleotide of base type Guanine (G) or a sequence with at least 90 % similarity thereto, wherein each base type X independently of each other designate base type Thymine (T) or base type Uracil (U). The nucleic acid probe is immobilized to the solid support using UV light. Further disclosed is a test system comprising a solid support having at least one supercritical angle fluorescence structure (SAF structure).