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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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Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

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Fluorometric RdRp assay with self-priming RNA.

Fatih Kocabas¹, Raife D Turan, Galip S Aslan

¹Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, 34755, Istanbul, Turkey, fatih.kocabas@yeditepe.edu.tr.

|March 10, 2015

Summary

This study introduces a new fluorometric assay for discovering antiviral drugs. The assay specifically targets RNA-dependent RNA polymerase (RdRp) in RNA viruses, offering a non-radioactive method for drug screening.

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Last Updated: Apr 16, 2026

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Published on: October 31, 2014

Area of Science:

Biochemistry
Virology
Drug Discovery

Background:

Current antivirals often lack specificity, leading to off-target effects.
There is a critical need for novel antiviral compounds with improved specificity.
Existing methods for identifying RNA-dependent RNA polymerase (RdRp) inhibitors frequently use radioactive assays.

Purpose of the Study:

To develop a novel, non-radioactive fluorometric assay for screening antiviral compounds.
To specifically target the RNA-dependent RNA polymerase (RdRp) of RNA viruses.
To establish a reliable method for assessing RdRp activity in vitro for drug discovery.

Main Methods:

Utilized DNA- and RNA-specific fluorophores for detection.
Developed a fluorometric approach to measure RdRp activity.
Employed a self-priming RNA strategy to assess RdRp function.

Main Results:

Successfully differentiated between double-stranded RNA and single-stranded RNA using fluorescence.
Established an optimized fluorometric assay for RdRp activity.
Demonstrated the utility of the assay for antiviral drug screening.

Conclusions:

The developed fluorometric assay provides an effective and specific method for identifying RdRp inhibitors.
This approach offers a valuable alternative to radioactive methods in antiviral drug discovery.
The assay is suitable for screening compounds targeting RNA viruses by inhibiting RdRp.