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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Updated: Jun 19, 2026

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition

Published on: February 9, 2024

Efficient nucleic acid detection by templated reductive quencher release.

Raphael M Franzini¹, Eric T Kool

¹Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.

Journal of the American Chemical Society

|November 6, 2009

Summary

This summary is machine-generated.

New Q-STAR probes enable rapid, sensitive, and specific nucleic acid detection. This fluorescence activation strategy allows visual identification of genetic information, even distinguishing bacterial species by single nucleotide differences.

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Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

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Published on: September 24, 2015

Area of Science:

Biochemistry
Molecular Biology
Analytical Chemistry

Background:

Nucleic acid detection is crucial for diagnostics and research.
Current methods can be time-consuming or lack sensitivity.
Direct visual detection in living cells remains a challenge.

Purpose of the Study:

To develop a novel RNA-templated fluorescence activation strategy.
To create highly sensitive and specific DNA probes for nucleic acid detection.
To enable direct visual detection of genetic material in vitro and in cells.

Main Methods:

Design and synthesis of "Q-STAR" (Quenching-release by StAARting a Reduction) probes.
Utilizing a phosphine-induced reduction of an azide-containing linker for fluorescence activation.
Demonstrating probe performance using bacterial 16S rRNA and single nucleotide polymorphism discrimination.

Main Results:

Q-STAR probes achieved strong fluorescence turn-on signals within 20 minutes.
The system exhibited very low background noise and significant signal amplification.
A green/red probe pair successfully differentiated bacterial species based on a single nucleotide difference in 16S rRNA.

Conclusions:

The reductive quencher release mechanism provides a robust method for fluorescence activation.
Q-STAR probes offer a promising platform for sensitive and specific nucleic acid detection.
This technology has broad potential for applications in molecular diagnostics and live-cell imaging.