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.
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Kudla, M., Karginov, F.V. (2016). Measuring mRNA Translation by Polysome Profiling. In: Lin, RJ. (eds) RNA-Protein Complexes and Interactions. Methods in Molecular Biology, vol 1421. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3591-8_11
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DOI: https://doi.org/10.1007/978-1-4939-3591-8_11
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3591-8
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