Protocols > Arabidopsis thaliana Thylakoid Preparation

Protocol

1. Harvest 2-5 g leaves and place in a 50 ml Falcon tube containing 25 ml of cold Homogenization buffer.
2. Homogenize using an ultra-turrax (5 x 3 seconds).
3. Place 3 layers of cheesecloth on top of a 20 μm nylon mesh in a funnel.
4. Filter homogenized mix into a 40 ml centrifuge tube.
5. Centrifuge at 6500 rpm (5000 g) for 2 min in JA20 rotor.
6. Resuspend pellet in 25 ml of Homogenization buffer.
7. Homogenize further using a glass homogenizer.
8. Centrifuge at 6500 rpm (5000 g) for 2 min in JA20 rotor.
9. Resuspend pellet in 25 ml of Resuspension buffer.
10. Homogenize further using a glass homogenizer.
11. Centrifuge at 6500 rpm (5000 g) for 2 min in JA20 rotor.
12. Repeat steps 9 to 11 once or twice.
13. Resuspend in 1 ml of Storage buffer, more or less depending on color.
14. Store at -20°C.

Solutions