Keep in mind that following sterile procedures is crucial in mushroom cultivation to avoid contamination. Here’s a basic Standard Operating Procedure (SOP) for mushroom spore inoculation on agar, in grain bags, and using brown rice flour (BRF) cakes:

Agar Inoculation:

Materials Needed:

1. Agar plates with your chosen medium

2. Mushroom spore syringe

3. Sterile gloves

4. Flame source (alcohol burner or torch)

5. Sterile scalpel or inoculation loop

6. Sterilized work area (flow hood or still air box)

Procedure:

1. Sterilize your work area and all materials using a flame.

2. Put on sterile gloves.

3. Flame the scalpel or inoculation loop until it’s red hot.

4. Use the sterilized tool to open the spore syringe.

5. Flame the mouth of the spore syringe.

6. Insert the syringe needle into the agar, lifting the lid slightly to create an opening.

7. Inject a small amount of spores onto the agar surface.

8. Close the lid and label the plate with the strain and date.

9. Incubate the agar plate in a clean, dark area at the appropriate temperature.

Grain Bag Inoculation:

Materials Needed:

1. Sterilized grain bags

2. Mushroom spore syringe

3. Sterile gloves

4. Flame source

5. Alcohol wipes

Procedure:

1. Sterilize your work area and all materials.

2. Put on sterile gloves.

3. Flame the needle of the spore syringe.

4. Inject a small amount of spores into the grain bag through the injection port.

5. Seal the injection port with a sterile piece of tape.

6. Mix the spores throughout the grain substrate by gently shaking or massaging the bag.

7. Allow the mycelium to colonize the substrate in a clean, dark area at the appropriate temperature.

BRF Cake Inoculation:

Materials Needed:

1. Sterilized jars with BRF substrate

2. Mushroom spore syringe

3. Sterile gloves

4. Flame source

5. Alcohol wipes

Procedure:

1. Sterilize your work area and all materials.

2. Put on sterile gloves.

3. Flame the needle of the spore syringe.

4. Inject a small amount of spores into each jar through the inoculation holes.

5. Seal the inoculation holes with sterile tape.

6. Gently shake or swirl the jars to distribute the spores in the substrate.

7. Incubate the jars in a clean, dark area at the appropriate temperature until fully colonized.

8. Once colonized, move on to the fruiting stage.