Aqief Afzal, ASRBC, ACI Seed, ACI Ltd:The first and foremost rule is to build up or construct a well-organized lab. For this, some rules must be maintained very strictly.
• The working room, media preparation room, washing room and the incubation room must be separated from one other. In no case the rooms can be left open from any side and separate doors must be maintained to separate one from the other.
• The washing zone must be kept at a distance from the working room. Or there will be a chance of contamination as spores are always moving freely in the air. If possible air curtains are to be used in front of the door of the working room and incubation room.
• All the instruments used for the tissue culture lab must be separate from the others and also separate autoclave machine is to be used.
• Separate lab coats and sandals to be used for the lab as well and those are not to be brought outside of the working room.
• The ambient temperature must always be maintained in the growth room for uniform growth. Also providing the required amount of light for required amount of time is a must.
Preventing Contamination in the Tissue Culture Lab:
There are times when contamination becomes a major issue which creates a loss in the overall plantlet production procedures. If the following points are followed then the contamination rate can be brought down.
• The most frequent time of contamination is during the rainy season. This happens if the walls somehow become damp. This cannot happen at any cost.
• The glass vessel used for growing plantlets must be thoroughly cleaned and autoclaved. There is no alternative to this.
• The contaminated vessels cannot be opened normally. At first, the vessels are to be autoclaved and then thoroughly washed. Then it is to be autoclaved again before using. After media preparation, the vessels are to be autoclaved again.
• Only authorized personnel and person related to the work must enter the working room and the growth rooms. This should be strictly followed.
• The working hood or the laminar hood must not be very close to the door.
• Always using sterile instruments is a must.
• Fumigation is to be given after every 2 months. When contamination rises fumigation cave be given every alternate month.
• There are times the pH in the media also becomes a factor for the increased rate of contamination. The pH level must be strictly maintained for the specific plantlets.
If the above-mentioned points are strictly followed and practiced then the rate of contamination will be less.