AAT Bioquest

What methods can I use to measure and detect translation?

Posted February 24, 2023


There are several different methods one can use to measure and detect translation of mRNA. One conventional technique is the genetically tagged pulse-chase approach, in which newly synthesized proteins are uniquely tagged. In this method, the fluorescence tags are inserted into target genes, which create very specific fluorescence signals. Some novel techniques have been made for genetically encoded proteins to bind to different ligands. This method helps to localize newly synthesized proteins and for the identification of translation-rich regions. Fluorescence assays can also be used to detect ribosome interactions with mRNA, such as using FISH to detect ribosomes that are located close to the target mRNAs.

Another method is non-genetic imaging techniques, which allows for the visualization of the translation of endogenous mRNA. These techniques are used to study the distribution of translation sites and newly synthesized proteins. Isotopically labeled amino acids are one type of non genetic imaging technique. They use very specific vibrational energy of a carbon-deuterium bond that is able to be measured using Raman scattering microscopy. 

More novel techniques have also been developed such as using a biosensor to measure translational characteristics of single mRNA in live cells. The biosensor is used to directly visualize the first steps of translation using multicolor RNA labeling. NGS techniques such as polysome profiling, Ribo-Seq, and Trap-Seq can also be used. Polysomal profiling is used via micro-arrays or RNA-seq based quantification of polysome correlated mRNAS. It provides reproducible, and high quality analysis, as well as instant visualization.

Additional resources

Translatomics: The Global View of Translation

ReadiUse™ TCA Deproteinization Sample Preparation Kit