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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

3.8K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.8K

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Related Experiment Video

Updated: Nov 1, 2025

Polysome Profiling without Gradient Makers or Fractionation Systems
05:56

Polysome Profiling without Gradient Makers or Fractionation Systems

Published on: June 1, 2021

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Polysome Profiling without Gradient Makers or Fractionation Systems.

Mack Sobhany1, Robin E Stanley2

  • 1Signal Transduction Laboratory, National Institute of Environmental Health Sciences, Department of Health and Human Services, National Institutes of Health; sobhany@niehs.nih.gov.

Journal of Visualized Experiments : Jove
|June 21, 2021
PubMed
Summary
This summary is machine-generated.

This study presents an accessible protocol for generating ribosome profiles using standard molecular biology equipment, avoiding costly specialized fractionation systems. The method ensures reproducible polysome profiles for research applications.

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Last Updated: Nov 1, 2025

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Polysome fractionation via sucrose density gradient centrifugation is crucial for analyzing mRNA translation and ribosome-associated factors.
  • Specialized automated systems for this technique are often expensive, limiting accessibility for many research labs.

Purpose of the Study:

  • To present a cost-effective protocol for reproducible polysome profiling using standard laboratory equipment.
  • To compare polysome profiles generated with and without specialized gradient fractionation systems.

Main Methods:

  • Sucrose density gradient centrifugation was performed using standard equipment.
  • Ribosome profiles were generated and analyzed for Saccharomyces cerevisiae.
  • A comparison was made between profiles obtained with and without automated fractionation systems.

Main Results:

  • Reproducible polysome profiles were successfully generated without specialized fractionation instruments.
  • The protocol demonstrated adaptability for various organisms and cell types.
  • Optimization strategies for reproducible profiling were discussed.

Conclusions:

  • A cost-effective and accessible method for polysome profiling is established.
  • This protocol empowers researchers with limited resources to study mRNA translation.
  • The findings facilitate broader research into translational regulation across different biological systems.