Another methodology to inoculate your grain, is by first propagating the mushroom tissue on Agar (or cloning). Measure out 5.75 grams of nutrient agar powder to 1 cup of unpolluted water (ample for five or extra Petri dishes). Begin to warmth and stir till the agar is totally dissolved. As it begins to boil, proceed to stir for a minute after which take away from the warmth. Pour a skinny layer into your Petri dishes and canopy with lids. Wrap in aluminium foil and stress cook dinner together with your grain (or for at the least 30 minutes at 15 psi). Move your Petri dishes, mushroom tissue and different gear to the clear room. Allow the Petri dishes to chill fully. Spray the clear room partitions, benches and flooring with 5% bleach answer (as earlier than put on clear garments, wash your palms and many others). A laminar movement bench and hepa filter would cut back contamination throughout this stage, however it’s potential (with a better contamination charge) to succeed with out one.
Taking the mushroom by its base, rigorously spit it in two. Place the mushroom (exterior down) on to a clear floor, ensuring you retain the within tissue from touching something. Sterilise the scalpel blade by holding it throughout the alcohol burner’s flame. Lift the lid of the Petri dish and funky the scalpel blade by putting it centrally into your agar. With the scalpel, rigorously reduce a small sq. from the newly uncovered mushroom tissue. Place the sq. of tissue centrally into the agar and canopy with the Petri dish lid. You might want to tape the lid (with a clear breathable tape) to cut back the possibility of contamination. Repeat the method, ensuring to sterilise the scalpel earlier than every switch. Leave the Petri dishes to incubate.
Incubate for pleurotus ostreatus at 24°C (75°F) and for pleurotus pulmonarius (summer time) 24°C to 30°C (75°F to 85°F). Colonisation ought to take roughly eight to 10 days.
During this time, take away any Petri dishes that seems to be contaminated with different moulds. Once totally colonised (mycelium nearing the sides), it’s time to switch the agar to the sterilised grain. Choose solely probably the most wholesome cultures for the inoculations. Sterilise the scalpel blade, take away the Petri dish lid and reduce 2 wedges from the centre of the dish. Remove the aluminium foil and lid of the grain jar. Using the scalpel, switch the wedges to the sterile grain. Quickly, push a small quantity of cotton wool by means of the lid’s respiration gap and fasten to the jar. Finally, shake the jar vigorously to disperse the mycelium from the agar all through the grain. Place on a shaded shelf throughout the clear room to incubate (temperatures as above). Repeat for every jar of sterilised grain.
Note: Cloning repeatedly (with out introducing new strains) might result in replicate fading, with subsequent cultures ultimately shedding vitality and due to this fact producing much less mushrooms. In distinction to cloning, spores when germinated (see Step 8b), create many various strains that compete with one another. The ensuing mushroom traits might due to this fact differ from tradition to tradition.