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Related Concept Videos

Labeling DNA Probes03:31

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...
DNA Microarrays02:34

DNA Microarrays

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...
RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Related Experiment Video

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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
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Published on: December 21, 2017

Imaging individual mRNA molecules using multiple singly labeled probes.

Arjun Raj1, Patrick van den Bogaard, Scott A Rifkin

  • 1Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. arjunraj@mit.edu

Nature Methods
|September 23, 2008
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method to visualize individual messenger RNA (mRNA) molecules in fixed cells using fluorescent probes. This technique allows for precise identification and counting of specific mRNA species, enabling deeper insights into gene expression.

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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

Published on: August 6, 2014

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Accurate visualization of individual mRNA molecules is crucial for understanding gene expression dynamics.
  • Existing methods often lack the resolution or specificity to quantify single mRNA transcripts within cells.

Purpose of the Study:

  • To develop and validate a novel method for high-resolution imaging of individual mRNA molecules in fixed biological samples.
  • To enable simultaneous detection and quantification of multiple mRNA species within single cells.

Main Methods:

  • Utilizing a large number (48+) of short, singly labeled oligonucleotide probes to hybridize with each target mRNA species.
  • Employing fluorescence microscopy to detect the fluorescent spots generated by probe binding.
  • Developing computational algorithms for identifying and localizing individual fluorescent spots corresponding to mRNA molecules.

Main Results:

  • Demonstrated the ability to visualize individual mRNA molecules as computationally identifiable fluorescent spots.
  • Successfully achieved simultaneous detection of three distinct mRNA species in single cells.
  • Validated the method's applicability across diverse biological systems, including yeast, nematodes, fruit fly tissues, and mammalian cells/neurons.

Conclusions:

  • The developed probe-based imaging method provides a powerful tool for single-molecule mRNA detection.
  • This technique offers high specificity and sensitivity for analyzing mRNA expression patterns in various organisms and cell types.
  • Enables quantitative analysis of gene expression at the single-cell level, advancing molecular and cell biology research.