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Protocol should be developed first.
Glasswares/Distilled water and forceps,needle,Blade,scissors etc.must be autoclaved in proper containers/Al foils and kept under Laminar Air flow Cabinet.
LAF working area must be sterilised with UV light and rectified spirits etc before use.
Explants must be sterilised with proper disinfectants and sterile water before transfer under HEPA filter Air flow.
Proper Medium must be prepared as per requirement and autoclaved. Should be used after3-4 days to see infections,if any.
Inoculation must be made and glasswares/bottles must be sealed under sterilised conditions.
Glasswares must be transferred to the culture Room with controlled conditions.
Further steps -sub culturing etc.must be carried out as per protocol.
It is difficult to handle as first of all the laboratory or working area should be sterilized by UV light. After that, culture plates, media and flasks should be autoclaved before using them for tissue culture. The plant tissue or explant must be sterilized. Lastly, all the work should be done according to the protocol in laminar flow hood. Laminar flow hood should be cleaned with 70% ethanol.
it's realy sort of dificult because you have to first sterilize the laboratory and the use the ultra violet rays to reduce contamination and use autoclave for the media or ultra filtration for the media that you can't use autoclave with, the last thing is you must be sure that your tissues are contamination free first
