Journal: Plant Methods

Article Title: Protocol: Optimised methodology for isolation of nuclei from leaves of species in the Solanaceae and Rosaceae families

doi: 10.1186/1746-4811-9-31

Figure Lengend Snippet: Preparations of nuclei obtained following the final purification on 35% Percoll cushion by centrifugation. Preparations of nuclei from leaves of (A) tobacco, (B) potato and (C) apple obtained after the final purification on 35% Percoll cushion by centrifugation (step 6 in Figure A) contained only negligible amounts of debris. DAPI-stained nuclei (shown in blue) were observed with a fluorescence microscope using UV-light and debris observed simultaneously under normal light. The unstained structures are remaining contaminants. Scale bars, 10 μm.

Article Snippet: Blades Cheesecloth Falcon tubes (15 ml, 50 ml) Funnels Glass container Homogenizer (UltraTurrax T25, IKA) Hybond-P polyvinylidene fluoride membrane (GE Healthcare) Leitz Laborlux S microscope with an epifluorescence extension (Leitz Ploemopak) and DAPI filter Miracloth Mortar and pestle Orbital shaker incubator Pasteur pipettes Swinging rotor centrifuge ( e . g ., Eppendorf 5804R)

Techniques: Purification, Centrifugation, Staining, Fluorescence, Microscopy