- Protoplasts are filtered through a nylon mesh (64micrometer)
to remove undigested tissue, cell clumps, and cell wall debris.
- Transfer filtrate to centrifuge tube and spin at +
75 x g (5 min).
- Debris (in supernatant) is carefully removed (protoplasts
have formed a pellet at the base of the tube).
- Protoplasts are carefully resuspended in culture medium
(plus 13% mannitol), and the process is repeated three times.
- Protoplasts are examined for density and viability.
Nylon mesh filter
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