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Select well grown, disease free oyster mushroom early in the morning and keep it on a clean paper for 2-3 hr. to get certain amount of moisture present in the mushroom to get evaporated.
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Clean the culture room/ laminar flow chamber with antiseptic solution.
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Keep the sterilized PDA slants, razor blades, forceps etc. inside the chamber and put on the UV light.
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After 20 minutes put off the UV light and and start working after 5 minutes.
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Sterilize all the instruments to be used by exposing to Bunsen burner.
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Surface sterilize the mushroom with ethyl alcohol using absorbent cotton and split open the mushroom longitudinally into two halves.
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Using a new, sterilized blade cut a small piece of tissue from the centre of the split mushroom at the junction of pileus and stipe.
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Remove the cotton plug of the agar slant and the tissue is aseptically placed inside the slant by using a sterilized forceps and close it immediately.
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After transferring tissues from the mushroom, the tube are arranged in a wire basket and kept in a clean room at room temperature for the growth of the fungus.
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Observe the tube at periodical intervals and remove the contaminated ones. The tubes will be ready for further use within another ten days. The culture is used for preparation of mother spawns.
