[Baseclick Newsletter] Click Chemistry in Fluorescently Labeled Oligonucleotides for qPCR_BaseClick - 코아사이언스

안녕하세요!

Baseclick 수입/공급 업체 코아사이언스 입니다.

 

자세한 상품 정보 및 관련 문의는 전화 02-858-0328 또는 info@coresciences.co.kr로 문의주시기 바랍니다.

감사합니다!

Click Chemistry in Fluorescently Labeled Oligonucleotides for qPCR

Click chemistry has revolutionized nucleic acid modification, providing a highly efficient and selective method for attaching functional groups such as fluorophores and quenchers to oligonucleotides. In particular, the copper-catalyzed azide-alkyne cycloaddition (CuAAC) enables site-specific modifications with high yield and minimal side reactions. This is crucial for applications like quantitative PCR (qPCR), where precise fluorescence labeling ensures accurate detection and quantification of target sequences.

 

Applications of Click Chemistry in qPCR

1. Fluorescent Oligonucleotides as Beacons for qPCR

Molecular beacons (MBs) are single-stranded oligonucleotide probes that form a hairpin structure, keeping the fluorophore and quencher inclose proximity until hybridization occurs. Upon binding to a target sequence, the structure unfolds, allowing fluorescence emission. This approach provides:
• High specificity due to precise hybridization with complementary sequences.
• Low background signal when the probe is not bound to the target.
• Real-time monitoring for accurate quantification in qPCR.
• High-efficiency labeling without compromising the integrity of the oligonucleotide structure.
• Multiplexing capabilities, enabling simultaneous detection of multiple targets using different fluorophores.

Recent advancements in beacon chemistry and dye selection have improved thermal stability and signal-to-noise ratio, making them even more reliable for clinical and research applications.

2. Improved Stability of Probes
Traditional labeling methods often involve reactive dyes that can degrade or compromise oligonucleotide stability. Click-modified oligonucleotides offer:
• Enhanced chemical and thermal stability, improving the shelf life and reproducibility of qPCR assays.
• Better performance in high-temperature reactions, crucial for reliable qPCR results.

3. Custom Probe Development for Molecular Diagnostics
Click chemistry facilitates the development of dual-labeled probes with optimized fluorescence resonance energy transfer (FRET) properties by enabling efficient quencher integration.

This is particularly useful for:
• Clinical diagnostics, including pathogen detection (e.g., COVID-19, HPV) and cancer biomarker analysis.
• Personalized medicine, allowing for customized probe development tailored to specific genetic targets.

4. On-Demand Synthesis Using Enzymatic DNA Synthesis (EDS)
Our partner DNA Script leverages click chemistry for enzymatic synthesis of modified oligonucleotides.

DNA Script utilizes click chemistry to enable the modification of oligonucleotides with various functional groups, such as dyes, quenchers, and other biologically relevant molecules. DNA Script's SYNTAX™ DNA printing system allows for the enzymatic synthesis of high-purity nucleic acids with these modifications, providing a rapid and flexible solution for creating customized oligos for research and diagnostic applications.
• Empowers researchers to generate customized qPCR probes in their own laboratories.
• Reduces reliance on third-party synthesis services, improving flexibility and turn around time.

 

 

Advantages of Click Chemistry in Fluorescent Oligonucleotide Labeling

High Specificity & Efficiency:

Enables site-specific modifications without interfering with nucleic acid function.
Mild Reaction Conditions:

Operates under biocompatible, non-damaging conditions, unlike traditional chemical labeling methods.
Versatile Functionalization: Allows incorporation of fluorophores, quenchers, biotin, PEG modifications, and other functional groups.
Fast & Scalable:

The reaction is rapid and highly efficient, making it suitable for high-throughput probe production.
Eco-Friendly Alternative:

Minimizes waste and avoids toxic by-products associated with conventional labeling techniques.

Future Perspectives
The integration of click chemistry with fluorescent oligonucleotide synthesis paving the way for more sensitive and efficient diagnostic tools.

Future applications are expected to expand significantly in:
• Infectious disease detection (e.g., emerging viral threats).
• Cancer diagnostics (e.g., mutation-specific probes).
• Personalized medicine, enabling patient-specific genetic screening.
Continued innovation in click chemistry, enzymatic synthesis, and probe design will further enhance the accuracy and reliability of qPCR-based assays.

 

코아사이언스 coresciences baseclick korea distributor 한국 대리점 Oligonucleotides Surface Engineering Tools PEG and Linkers Non Fluorescent Tags Fluorescent Tags Click Reagents Free Nucleosides Peptide Pools Peptide Pools Amino Acid Labeling Modified and Standard Triphosphates RNA Labeling FISH Kits & Probes Metabolic Labeling Transfection Agents Cell Stains and Imaging Reagents Cell Proliferation Kits 올리고핵산 올리고뉴클레오티드 표지