Protocol for Quantifying Silent Ribosome Induction Using Polysome Profiling

Translationally silent ribosomes have become an important focus in RNA biology. Lacking mRNAs, they represent a hibernating ribosome state. Their quantification remains challenging. This protocol outlines how to measure silent ribosome induction under diverse conditions by analyzing RNA from polysome-profiling fractions in yeast and mammalian cells, using RNA-seq with detection and removal of anomalously amplified rRNAs, complemented by qPCR for validation. For an example of the execution of this protocol, please refer to Rahaman, et. al [1].