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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: Feb 28, 2026

Polysome Profiling without Gradient Makers or Fractionation Systems

Polysome Profiling without Gradient Makers or Fractionation Systems

Published on: June 1, 2021

Polysome Analysis.

Dipak Kumar Poria¹, Partho Sarothi Ray¹

¹Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India.

|June 13, 2017

Summary

This summary is machine-generated.

Polysome analysis separates actively translating messenger RNAs (mRNAs) from non-translating ones using sucrose density gradient ultracentrifugation. This method helps investigate mRNA translational states by detecting mRNA association with ribosomal fractions.

Keywords:

Polysome Ribosome fractionation Sucrose density gradient Translation Ultracentrifugation mRNA

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Last Updated: Feb 28, 2026

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
Biochemistry
Genetics

Background:

Understanding mRNA's translational state is crucial for gene expression regulation.
Polysome analysis provides insights into which mRNAs are actively translated within a cell.

Purpose of the Study:

To describe a protocol for polysome analysis.
To differentiate between actively translating and non-translating mRNAs.

Main Methods:

Cell lysates are fractionated using sucrose density gradient ultracentrifugation.
Fractions containing free mRNA, 40S, 60S ribosomal subunits, monosomes, and polysomes are collected.
Reverse transcription followed by PCR (RT-PCR) is used to detect specific mRNA presence in each fraction.

Main Results:

The protocol successfully separates various mRNA populations based on their translational activity.
Differential association of mRNAs with ribosomal fractions indicates their translational status.

Conclusions:

Polysome analysis is an effective method for assessing mRNA translational states.
This technique aids in studying the regulation of gene expression at the translational level.