• Techniques Of Tissue Culture

Schematic Diagram of techniques of 
Tissue Culture

Steps of Tissue Culture

1. Surface sterilization: 

The explant ( the plant plant part excised for the in-vitro cultivation) is surface sterilized to eliminate contaminants using sodium hypochlorite @1-2 per cent or mercuric chloride@ 0.1% per cent solution.

2. Nutrient Medium

It is a medium containing salts, trace elements, vitamins, carbon sources, growth regulators 2,4-D @ 0.5-0.2 mg/L is commonly used. Organic supplements like coconut agent such as agar-agar is used. generally B-5 medium (Gamborg et al.)or MS medium (Murashige and Skoog )are used.

3. Sub Culturing

Transferring of tissue to fresh media after a stipulated time. Usually sub-culturing is done after every 4-6 weeks. However, the suspension cultures are sub-cultured after every 3-14 days.

4. Plant Regeneration and transfer         to soil

First the transfer of cultured plant is done in small pots and then to greenhouse and finally in soil. Transfer is done when roots and soots appears.