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Caution : Take care with powdered calcium hypochlorite as it is a powerful reducing agent. If calcium hypochlorite is stored moist and the container opened later, it can explode. Store calcium hypochlorite in a sealed container in a dry place.

A summary of the six steps commonly involved in establishing and maintaining aseptic plant tissue culture follows.

i.  Collect pieces of plant material (ex-plants) in a screw-cap bottle. Immerse them in a dilute solution of the disinfectant containing a wetting agent. Replace the lid and store the bottle in the laminar air flow cabinet. Shake the bottle two or three times during the sterilization period.

ii.   Remove the lid and drain carefully. Thoroughly rinse the plant material in sterilized distilled water and replace the lid. After shaking a few minutes, discard the water. Rinse two or three times more.

iii.   Transfer the material to a pre-sterilized Petri-dishes or test-tubes.

iv.  Sterilize the required instruments by dipping them in 70% ethanol and flamed them. Allow to cool. Sterilize the instruments after each time they are used to handle tissue.

v.  Prepare suitable explants from the surface sterilized material using sterilized instruments (scalpels, needles, forceps, etc.).

vi.  Quickly remove the lid of the culture vessel, transfer the explants on to the medium, flame the neck of the vessel (only if glass) and replace the lid.

If handling aseptic plant materials during routine subculture, omit the first two steps.