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INOCULATION
It has the laminar Air Flow cabinet, which generates clean air, with HEPA filter of pore size 0.2 mm. Bacteria and fungal spores, higher than 0.2 mm get killed and cannot pass through these filters. Excised explants are surface sterilized with 0.1% Mercuric chloride solution for 10 minutes, after three washings with autoclaved distilled water. The outer leaves are asceptically removed to obtain suitable spindle explants (0.5 – 1.0 cm). Individual explants are inoculated/cultured on MS (1962) establishment media.
MULTIPLICATION
The established cultures are then multiplied on multiplication media by re culturing and sub culturing these cultures after cleaning on fresh liquid media. These are sub cultured by changing the media after every 15 days.
INCUBATION
After marking the culture bottles for different varieties, cultures are incubated in the incubation room with a specified temperature between 25 ± 2 Degrees Centigrade which is maintained with the help of air conditioners, as at lower temperature, growth slows down and higher temperature (³ 30 Deg. Centigrade) results in the death of the plants. Light is another important factor; cultures are incubated on the culture racks fitted with fluorescent tube light rods to meet requirements of high intensity light (5000 Lux) for the growth of cultures. Duration of light exposure is kept between 16 hours light and 8 hours dark. High humidity (60-70%) is maintained in the growth room to avoid media dessication.
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