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Ribosome Profiling

Ribosome Profiling

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...

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Related Experiment Video

Updated: Mar 14, 2026

An Easy Method for Plant Polysome Profiling

An Easy Method for Plant Polysome Profiling

Published on: August 28, 2016

An Easy Method for Plant Polysome Profiling.

Cécile Lecampion¹, Maïna Floris², Jean Raphaël Fantino³

¹Laboratoire de Génétique et Biophysique des Plantes, Aix-Marseille Université; UMR 7265 Biologie Végétale & Microbiologie Environnementales, CNRS; BIAM, CEA; cecile.lecampion@univ-amu.fr.

Journal of Visualized Experiments : Jove

|September 30, 2016

Summary

This study presents an easy, economical method for polysome profiling in plants, crucial for analyzing translational regulation. The technique simplifies ribosome-mRNA complex fractionation for a snapshot of global translation activity.

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An Easy Method for Plant Polysome Profiling

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Polysome Profiling without Gradient Makers or Fractionation Systems

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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Area of Science:

Molecular Biology
Plant Science
Biochemistry

Background:

Translation of mRNA to protein is a fundamental biological process.
Polysome profiling is the gold standard for analyzing translational regulation.
Existing methods can be complex and costly.

Purpose of the Study:

To describe an easy and economical method for polysome profiling in various plant tissues.
To enable analysis of translational regulation in plants.
To provide a snapshot of global translational activity.

Main Methods:

A simplified sucrose gradient is created by sequential freezing.
Cytosolic extracts are prepared with ribosome inhibitors (cycloheximide and chloramphenicol).
Sucrose gradient ultracentrifugation followed by direct fraction collection and continuous A260nm monitoring.

Main Results:

Six fractions are collected, yielding distinct polysome, monosome, and unbound mRNA populations.
A polysome profile is generated, reflecting global translational activity.
The protocol is validated across multiple plant species and tissues (Arabidopsis, Nicotiana, Solanum, Oryza).

Conclusions:

This method offers an accessible and cost-effective approach to polysome profiling in plants.
It facilitates the study of translational regulation across diverse plant systems.
The protocol provides valuable insights into global translational activity in plants.