Inoculating Sterilized Mushroom Grain Spawn – All Methods Explained

In this guide, you will learn how to inoculate sterilized mushroom grain spawn using a liquid culture syringe, spore syringe, colonized spawn, or agar culture.

As a side note, if the genetics you are using (spores, liquid culture, agar culture) are contaminated, the sterile grain spawn will not work. Please ensure your genetics come from a reliable supplier and have been tested.

Cleanliness

Keeping your workspace, spawns, substrate, and cultures clean and sterile is essential to successful mushroom cultivation. Each step of the process takes a significant amount of time, and the last thing you want is for you to introduce competing bacteria to your developing mycelium, resulting in a contaminated grow.

Take the time to clean all of your tools, yourself, workspace, and supplies with isopropyl alcohol, as well as ensure spawns and substrates are sterilized correctly.

How to Inoculate Sterilized Mushroom Grain Spawn

There are several types of genetics you can use to inoculate your mushroom spawn, such as colonized spawn, agar tissue, spores, or liquid culture. 

Inoculating Grain Spawn Using a Syringe:

1. Clean the injection port area with isopropyl alcohol.
2. Shake your syringe well before injection.
3. Remove the syringe cap, and screw the needle into place.
4. Inject 5 ml to 10 ml of liquid solution through the injection port and into the bag.
5. Remove the needle and clean the injection port once more.
6. Place the bag in a warm, dark place for colonization. 

Needles are single-use only, unless you flame-sterilize.

Note: Colonization can take 2 to 6 weeks. Place the bag in a dark, warm spot (20C to 25C), and ensure the bag is not suffocated. Fresh air exchange will occur through the filter patch near the top of the bag.

Inoculating Grain Spawn Using Colonized Mycelium Spawn:

1. Clean both bags, especially near the tops where the bags will be open, with isopropyl alcohol.
2. With clean scissors, cut the seals at the top of both bags.
3. Pour a small amount of colonized spawn into the grain spawn bag via the top of the bag.
4. From the outside of the bag, mix the colonized spawn into the newly inoculated bag thoroughly.
5. Re-seal the top of both bags using an impulse sealer.
6. Place the bag in a warm, dark place for colonization. 

Note: Colonization can take 2 to 6 weeks. Place the bag in a dark, warm spot (20C to 25C), and ensure the bag is not suffocated. Fresh air exchange will occur through the filter patch near the top of the bag.

Inoculating Grain Spawn Using Agar Culture:

Any agar work should be done in front of a laminar flow hood or in a still air box. Performing agar tissue work in the open air will likely bring contamination.

1. With isopropyl alcohol. Clean the outside of the plate before removing the parafilm wrapping.
2. Flame-sterilize your scalpel blade and allow it to cool.
3. Cut 2 to 3 pieces of agar tissue from your plate, and transfer the tissue into the sterile grain spawn via the top of the bag.
4. From the outside of the bag, mix the agar tissue pieces with the grain spawn thoroughly.
5. Re-seal the top of the spawn bag with an impulse sealer and re-seal the agar plate with parafilm.
6. Place the bag in a warm, dark place for colonization. 

Note: Colonization can take 2 to 6 weeks. Place the bag in a dark, warm spot (20C to 25C), and ensure the bag is not suffocated. Fresh air exchange will occur through the filter patch near the top of the bag.

I hope this guide was helpful. If you have any further questions, please send us an email: info@flatlandfungi.com