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Related Concept Videos

Real Time RT-PCR02:57

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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Related Experiment Video

Updated: May 13, 2026

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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High-Throughput RT-PCR for small-molecule screening assays.

Joshua A Bittker1

  • 1Chemical Biology Platform, Broad Institute of MIT and Harvard, Cambridge, MA 617-714-7373.

Current Protocols in Chemical Biology
|March 15, 2013
PubMed
Summary
This summary is machine-generated.

Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) enables high-throughput screening of small molecules. This method analyzes gene expression changes, offering a powerful tool for drug discovery and development.

Keywords:
High Throughput ScreeningReal-time PCRgene expressionphenotypic screeningqRT-PCR

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Area of Science:

  • Molecular Biology
  • Pharmacology
  • Biotechnology

Background:

  • Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) is a standard for measuring mRNA expression levels.
  • Its application for assessing gene expression changes induced by small molecules or siRNA has been less frequent.
  • Advancements in instrumentation and commercial kits have paved the way for its use in high-throughput screening.

Purpose of the Study:

  • To detail the protocol for utilizing quantitative RT-PCR as a primary assay for high-throughput small-molecule screening.
  • To highlight specific considerations for employing RT-PCR in small-molecule screening applications.
  • To discuss methods for mRNA isolation and analysis within this screening context.

Main Methods:

  • Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR).
  • High-throughput screening assay development.
  • mRNA isolation and analysis techniques.

Main Results:

  • Quantitative RT-PCR can be adapted for high-throughput small-molecule screening, comparable to traditional HTS assays.
  • The protocol addresses essential considerations for this application.
  • Various mRNA isolation and analysis methods are suitable for this screening approach.

Conclusions:

  • Quantitative RT-PCR is a viable and scalable method for primary small-molecule screening.
  • This approach facilitates the analysis of gene expression changes in response to chemical compounds.
  • The protocol provides a framework for implementing RT-PCR in drug discovery pipelines.