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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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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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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Related Experiment Video

Updated: Mar 20, 2026

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

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IMAGEtags: Quantifying mRNA Transcription in Real Time with Multiaptamer Reporters.

J Ray1, I Shin2, M Ilgu3

  • 1Cornell University, Ithaca, NY, United States.

Methods in Enzymology
|June 1, 2016
PubMed
Summary
This summary is machine-generated.

IMAGEtag technology quantifies RNA levels in living cells, enabling researchers to track individual cell responses over time. This method allows dynamic monitoring of gene expression without cell sacrifice, advancing cell communication studies.

Keywords:
FRETFluorescenceIMAGEtagsRNA imagingYeast

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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biotechnology

Background:

  • Cellular communication is vital for multicellular organisms, relying heavily on dynamic RNA transcription.
  • Current methods for studying transcription often require cell sacrifice, limiting real-time analysis.
  • A need exists for technologies that allow continuous monitoring of gene expression in living cells.

Purpose of the Study:

  • To introduce and demonstrate the utility of IMAGEtag technology for quantifying RNA levels in living cells.
  • To enable longitudinal tracking of individual cell responses to environmental cues.
  • To provide a non-invasive method for studying dynamic gene expression.

Main Methods:

  • IMAGEtags are short RNA molecules with tandem aptamer sequences that bind specific small-molecule ligands.
  • Ligands are conjugated with fluorophores, forming Förster Resonance Energy Transfer (FRET) pairs.
  • Expression of an RNA containing an IMAGEtag sequence allows aptamer-ligand binding, bringing FRET pairs into proximity for detection.

Main Results:

  • IMAGEtag technology successfully quantified RNA levels in living yeast cells.
  • The system demonstrated sensitivity to report on messenger RNA (mRNA) expression levels.
  • Minimal background fluorescence in the FRET channel enhances signal detection.

Conclusions:

  • IMAGEtag technology offers a sensitive, non-invasive method for monitoring mRNA levels in real-time within living cells.
  • This technology facilitates the study of dynamic cellular processes and responses to stimuli.
  • IMAGEtagging opens new avenues for understanding cell communication and gene regulation.