Measuring mRNA Translation by Polysome Profiling - PubMed

Measuring mRNA Translation by Polysome Profiling

Marek Kudla et al. Methods Mol Biol. 2016.

Abstract

Determination of mRNA translation rates is essential to understanding the regulatory pathways governing eukaryotic gene expression. In this chapter, we present a transcriptome-wide method to assess translation by association of mRNAs with polysomes on sucrose density gradients. After sedimentation, the fractions are spiked with a control RNA mixture and the RNA content is measured by high-throughput sequencing. Normalization to the spike-ins provides a global quantitative view on the translational status of cellular mRNAs, with the ability to measure changes and identify active and silent subpopulations of each.

Keywords: Polysome; Sucrose gradient; Translation; mRNA.

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