Purification of Isolated Protoplasts - The protoplasts isolated as above are present in the media together with a range of cell debris and broken cell organelles. A number of methods are available for purification of protoplasts from this mixture. Only two commonly used methods will be briefly described in this section.

Sedimentation and washing:In this method, the crude protoplast suspension is centrifuged at low speed (50-100 g / 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 protoplst preparation.