How to make your own mushroom liquid culture?

6 min readMar 2, 2021


Mushrooms are mysterious organisms that are fleshy and spore-bearing in nature. They are the fruit of fungi (called Mycelium) that develop underground, inside trees, or rotting logs. Mushrooms grow from their spores. These mushroom spores are microscopic, yet they contain all the essential blueprints that mushrooms require for successful growth.

A mushroom spore develops into a solitary mushroom, and one Mushroom can create a huge number of spores. A fully developed mushroom is said to contain up to 16 billion spores. Spore prints are additionally being widely utilized for distinguishing various types of wild mushrooms.

Gourmet mushrooms

Gourmet mushrooms are earning increasing popularity across the globe. As shocking as it might sound, around one and a half million Gourmet mushrooms were grown in the United States alone. Oyster mushrooms, is likewise a type of gourmet mushrooms, have proven to be a hugely beneficial mushroom available. The best part about the Oyster mushrooms is that they can be grown quickly, in just six weeks. Hence, it’s conceivable to draw immense profits out of it pretty quickly.

This article would help you understand the whole process, which involves converting Gourmet Oyster Mushrooms into Liquid Mushroom Culture. It might sound complicated but, trust me, it’s really simple! Keeping up your mushroom culture collection is an efficient and remunerating experience. It requires some time to get its hang; however, there are techniques that you can use without spending too much money on buying expensive hardware.
You need to make sure that you do all the following steps in the order they are mentioned below. Before beginning, let me share some necessary information regarding liquid culture and its use in the whole procedure.

What Is a Liquid Culture?

Liquid culture, or LC as it’s regularly alluded to, is Mycelium suspended in a nutrient broth. The primary usage of LC is to make inoculating substrates simpler. This technique has more preference over agar culture since it helps ensure maximum sterilization. As the culture goes from a sterile zone (inside the jar) to another sterile environment using a sterile syringe, the chances of contamination are pretty low.

However, it should be noted that this technique has a few drawbacks, too, as it doesn’t develop on a leveled plane as agar does. Moreover, it’s essential to ensure that all the related equipment is not exposed to the encompassing contaminated environment; hence, great care should be taken while performing procedures involving Liquid Culture.

The easiest way to manufacture your own liquid culture is to buy a clean culture from a known supplier and then expand it into a more liquid culture. Add the glucose material into your distilled water inside the jar with an air filter, and then pressure cook the sweetened water for around 15–20 minutes at 15 PSI. The purpose of pressure cooking is to sterilize the water inside the jar. Leave it overnight, allowing it to cool. Once settled down, you can bring it to your lab and start working with it!

How to prepare Liquid Mushroom Culture using Oyster Mushrooms?

Put on your gloves

To begin with, put your gloves on. These would ensure that your hands are not a source of contamination in the working zone. Put sanitizer on your gloves for added safety.

Now, you are all set to begin this exciting procedure. It is recommended to perform this procedure in front of a fan or air filter (flow hood) to stabilize the environment.

Pick the “best” Mushroom

Foremost, you have to pick the Mushroom from your grow room or your farm. Remember a general rule while you do so: The bigger the stem, the more helpful Mushroom is for the procedure.

Disinfect your work zone

Next, clean up your working space. As mentioned before, your workplace must be free of all sources of contamination. Using a paper towel, clean all the visible dust or other residues from the exposed surface. Then, spray the surface adequately using Isopropyl Alcohol 70%. Once done, change your gloves, put on new ones, and sanitize again!

Sterilize a syringe and needle, and bring them over to the working table. Then, take your Liquid Culture Jar and spray Isopropyl Alcohol 70% on it.
Draw some of the liquid cultures into your sterilized syringe.

Next, take the needle, open it up and fix it into the syringe. Do it as close as possible to the Mushroom to avoid contamination, facing the fan. This ensures that the needle and the syringe remain in a sterile unit. Next, plunge the syringe into the jar with an air filter, which has been inoculated with sweetened distilled water. You have to draw some liquid out of the jar into the syringe. Once done, cover the needle immediately.

Extract the tissue from Mushroom (Needle biopsy)

Head towards your mushroom plant and pick one for yourself. Remember, choose the piece of Mushroom that has a more oversized stem, a firmer flesh.

Tear the Mushroom from the center, beginning from its cap, towards its stem. Face towards the fan while you do this. Keep one piece in your hand and the other on the table, ensuring that their sterile areas are exposed to the fan. (You could use the piece on the table as a backup plan.)
Here comes the central part. You have to plunge your syringe in the stem of the Mushroom in your hand, twist it a bit, making sure that it doesn’t leave from the other side. The purpose of this is to extract the flesh of the Mushroom. Gradually pull out the syringe. You would be able to observe the tiny piece of mushroom flesh on the needle’s tip now.

This whole procedure is referred to as needle biopsy and is often used in the medical field for diagnostic purposes.

Nice-to-know: Some people swab the Mushroom from its exterior surfaces after disinfecting it with alcohol instead of tearing it down and picking the piece of flesh from its center. This step isn’t wrong either; however, there are greater chances of contamination here as exposed regions are more prone to the encompassing polluted environment.

Inject the flesh into Liquid Culture

Next, you would directly take this tissue piece near your liquid culture and inject it into it. Now, you would be able to see the tiny tissue inside the liquid culture.

Stir your Liquid Culture

Next, you have to stir your culture. It is generally recommended to use a magnetic stir plate as it’s the most effective and safe method to do so. However, one can also add some marbles to the jar to do the deed.
The purpose of this stirring step is to ensure the availability of oxygen to the culture growing inside. It allows carbon dioxide to escape from the culture and oxygen into the culture, hence, making adequate oxygenation possible. Do it as frequently as you can. The better the stirring, the quicker would be your jar full of Mycelium. Mixing likewise helps break apart larger clusters of Mycelium, which boosts the culture growth rate.

How would you know if your Liquid Culture is free of contamination?

It’s really easy to deduce that. All you need to do is to put the liquid culture jar in front of a light source. The solution has to be clear. If it’s cloudy or milky, it implies that your culture has been contaminated.

Put it on a rack and wait for it to develop

Most of the mushrooms develop best at a temperature around 73F; they’ll grow a little slower outside of this. You should see the Mycelium gradually developing as it devours the sugar and starts respiring. Swirl your jar regularly to bring oxygen into the fluid and to break down larger pieces of Mycelium into smaller ones. The lesser their surface area, the more rapidly they would respire and hence, grow.

When the Mycelium starts taking up a lot of the container volume, energetically agitate the liquid culture until the Mycelium covers up the whole jar. Tilt it a bit, and allow some of the liquid to come against the top injection port. Inject a syringe, and suck some of the liquid cultures into it.
Your syringe is now ready to get injected into grain containers or move to agar as needed.


Despite the whole process is relatively simpler, it is significant to mention that there are chances of failure too. Every time we acquire a new skill, we don’t excel in it immediately. It takes time, patience, and consistency. Likewise is the case with mushroom growth. A little mistake from your side might ruin your whole procedure. Mostly, contamination during the entire working cycle is the main reason behind failures. Hence, ensure that you keep your working area sanitized, your clothes clean, and all your instruments sterilized while performing this complex microbiological procedure.

Repetitive practices and unwavering determination can undoubtedly help you make the perfect mushroom liquid culture, rich in Mycelium for yourself.

Let us know if you have questions about how to cultivate mushrooms or feel free to purchase our sterilized spawn bags in bulk. We will love to hear from you soon.

By Paul S.

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