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Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Author Spotlight: Advancements in DNA Nanosensors – Addressing Sensitivity and Selectivity Challenges in Molecular Detection

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Ligation-Based High-Performance Mimetic Enzyme Sensing Platform for Nucleic Acid Detection.

Xinrong Yan¹, Peiru Yang¹, Dehui Qiu¹

¹State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, P.R. China.

Analytical Chemistry

|December 28, 2023

Summary

This study introduces a novel chimeric peptide-DNAzyme (CPDzyme) integrated with ligase chain reaction (LCR) for highly sensitive nucleic acid detection. The LCR-CPDzyme system significantly enhances sensitivity for microRNA and single-nucleotide polymorphism detection.

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A Polyaniline-based Sensor of Nucleic Acids

A Polyaniline-based Sensor of Nucleic Acids

Published on: November 1, 2016

Area of Science:

Biochemistry
Molecular Biology
Nanotechnology

Background:

G-quadruplex (G4)/hemin DNAzymes offer potential as horseradish peroxidase substitutes in biosensing for nucleic acid detection.
Suboptimal catalytic activity of current G4/hemin DNAzymes hinders their application in highly sensitive biosensing platforms.
Development of improved signaling strategies is crucial for advancing nucleic acid detection sensitivity.

Purpose of the Study:

To develop a cost-efficient and highly sensitive method for nucleic acid detection using a novel chimeric peptide-DNAzyme (CPDzyme) integrated with ligase chain reaction (LCR).
To demonstrate the system's efficacy in detecting microRNA (miRNA) and single-nucleotide polymorphisms (SNPs).
To improve upon the sensitivity and accuracy of existing G4/hemin DNAzyme-based biosensing methods.

Main Methods:

Designed LCR probes with a G4-forming sequence and a terminal amino group.
Assembled asymmetric hemin with carboxylic arms, LCR products, and peptides to form CPDzyme.
Utilized magnetic separation for extraneous component removal followed by chemiluminescence detection.
Employed microRNA and single-nucleotide polymorphism detection as model systems.

Main Results:

The LCR-CPDzyme system achieved a 3-order of magnitude improvement in sensitivity compared to conventional G4/hemin methods.
Accurate quantification of miRNA down to 25 aM was achieved.
The system successfully differentiated 0.1% mutant DNA from a wild-type DNA pool.
Demonstrated enhanced catalytic capacity and improved signal generation.

Conclusions:

The proposed LCR-CPDzyme strategy offers a powerful approach for sensitive and accurate nucleic acid detection.
This method holds significant potential for in vitro diagnostics applications.
The strategy serves as a valuable reference for developing other ligation- or hybridization-based nucleic acid amplification assays.