Sedimentation and Washing, Crude Protoplasts Suspension, Centrifuged Low Speed, Intact Protoplasts, Pellet and Supernatant, Cell debris

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Protoplast Culture Regeneration and Somatic Hybridization Protoplast Culture Regeneration and Somatic Hybridization - Introduction Isolation of Proplasts Mechanical Method Enzymatic Method Direct One Step Method Sequential Two Step Method Purification of Isolated Protoplasts Sedimentation and Washing Flotation Viability and Plating Density of Protoplasts Testing Protoplast Viability Minimum Plating Density MPD and Culturing Individual Protaplasts Protaplast Culture and Regeneration of Plants Isolation of Subprotoplasts Protoplast Fusion and Somatic Hybridization Techniques of Protoplast Fusion

Home >> Plant Biotechnology and Genomics >> Protoplast Culture Regeneration and Somatic Hybridization >> Sedimentation and Washing

Sedimentation and washing

In this method, the crude protoplasts suspension is centrifuged at low speed (50-100g for 5 min). The intact protoplasts form a pellet and supernatant containing cell debris can be pipetted off.

The pellet is gently resuspended in fresh culture media plus mannitol and rewashed. This process is repeated two or three times to get relatively clean protoplast preparation.

Protoplasts can be purified by repeated gentle pelleting and resuspension

A. Crude Preparation	B. Impure Protoplast Pellet	C. Protoplast Suspension

F. Plating of Protoplasts	E. Resuspension	D. Purified Protoplast Pellet

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