Somatic Hybridization using Protoplasts
• Created by degrading the cell wall using enzymes
• Very fragile, can‟t pipette
• Protoplasts can be induced to fuse with one another:
– Electrofusion: A high frequency AC field is applied between 2 electrodes immersed in the suspension of protoplasts- this induces charges on the protoplasts and causes them to arrange themselves in lines between the electrodes. They are then subject to a high voltage discharge which causes them membranes to fuse where they are in contact.
– Polyethylene glycol (PEG): causes agglutination of many types of small particles, including protoplasts which fuse when centrifuged in its presence
– Addition of calcium ions at high pH values