Purification and analysis of chloroplast RNAs in Arabidopsis

Abstract

Chloroplasts are essential semiautonomous plant organelles responsible for photosynthesis, which generates sugars and oxygen vital for the entire biosphere. Additionally, chloroplasts regulate energy production, metabolite synthesis, and stress responses in plants and algae. Chloroplasts possess a notably complex RNA metabolism that includes RNA processing, editing, splicing, and regulation by various RNA-binding proteins. Highly purified chloroplasts, free of nuclear/cytoplasmic contaminants are desirable when studying chloroplast RNA metabolism. Here, we describe an efficient protocol to obtain highly purified chloroplasts for RNA analysis.