Rapid Miniprep of Intact Chloroplasts from <em>Arabidopsis thaliana</em> Leaves

Cell subfractionation is a common technique employed in many research laboratories to isolate organelles or intracellular compartments for the study of metabolism or biomolecule purification. While numerous protocols exist for isolating organelles, few are specifically designed for starting materials in the milligram range. Here, we present a detailed milligram-scale miniprep protocol for purifying intact chloroplasts from Arabidopsis thaliana leaves. This chloroplast miniprep procedure is suitable for applications such as confocal microscopy, western blotting, enzymatic assays, and other downstream analyses. Key features • This protocol condenses key aspects of existing methods and adapts them to the miniprep scale. • Researchers can isolate purified chloroplasts within 1.5 h. • This protocol requires standard laboratory equipment, such as microcentrifuges (ultracentrifuges are not required). Graphical overview.