Author:
Schiller Chiara,Reitter Sonja,Lehmann Janina A.,Fenzl Kai,Schott Johanna
Abstract
AbstractPolysome profiling is a powerful technique to study mRNA translation. After separation of ribosomal subunits from monosomes and polysomes by ultracentrifugation on sucrose density gradients, a UV absorbance profile is recorded during elution. This profile can be used to assess global translational activity, or reveal changes in ribosome biogenesis or translation elongation. In parallel to UV absorbance profiles, it is also possible to record fluorescence to measure the association of fluorescently tagged proteins with ribosomes or polysomes. To this end, the area under subsections of the UV/fluorescence profiles needs to be quantified carefully. In addition, alignment of profiles in one graph helps to visualize differences. With QuAPPro, we present the first interactive web app that allows quantification and alignment of polysome profiles, independently of the device or software that was used to generate the profiles. This user-friendly tool does not only speed up the analysis of polysome profiles but also facilitates reproducibility and documentation of the process.
Publisher
Cold Spring Harbor Laboratory
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