The Isolation of Total and Membrane-Bound Polysomes from Arabidopsis and the Detection of Their Associated AGO1 and sRNAs.

Polysomes play central roles in protein synthesis. Free polysomes are responsible for the translation of cytosolic and nuclear proteins, and the membrane-bound polysomes are responsible for the translation of transmembrane and secreted proteins. Accumulative evidence from recent studies shows that microRNAs are highly enriched on polysomes to regulate the expression of their own and target mRNAs. Here, we describe experimental protocols for the isolation of total and membrane-bound polysomes and the detection of the associated microRNAs and AGO1 proteins.