Making Liquid Culture Using Agar.
Making liquid culture from agar is a really simple process for those with experience in mushroom cultivation, however it does require a good understanding of sterile technique which can be difficult for novices.
Agar to liquid culture is one of the most widely used techniques for creating liquid culture.
It is preferred by mycologists due to its high success rate, as the mycelium on the agar plate is already fully colonized and healthy.
By using this method, the spawn has already been validated to a degree, and any potential contamination can be easily identified before use.
This is in contrast to using spores or other liquid culture to innoculate LC in which you cannot visibly see certain contamination until it has been tested.
In this post, we will go into detail about the process of inoculating liquid culture with agar, including the materials and equipment required, the benefits of using agar for liquid culture and any shortcomings with this technique.
Materials and Equipment
To ensure success and prevent contamination, it's important to have the proper materials and equipment on hand.
Here is a list of materials and equipment that are typically needed to make liquid culture from agar:
- Fully colonized agar plate of mycelium.
- Scalpel.
- Prepared liquid culture recipe (Karo, LME or Honey, each recipe is linked).
- Still Air Box.
- Alcohol.
- Gloves and face mask.
- Butane torch (get one of these for flaming your scalpel, makes it very easy).
- Pressure cooker or autoclave (if preparing liquid culture medium from scratch).
Step-By-Step Instructions On How To Make Liquid Culture From Agar.
To inoculate liquid culture with agar you will need a fully colonized agar plate and prepared liquid culture recipe. Once you have these you can begin the process.
Collect Items.
First thing you need to do is grab all the items you will need for inoculation and place them in your workspace.
Ensure you are working in an area you are comfortable which does not have any windows cracked open or heavy drafts.
Prepare Yourself And Work Space.
Put your face mask and rubber gloves on first then wipe them down with alcohol solution.
Using soapy water lightly wipe down your still air box. Don’t soak the still air box, instead lightly cover all the walls with a layer of soapy water.
Any contaminants which hit these walls will stick to the soapy water if there is air turbulence instead of moving around.
Put everything inside, close the lids and allow the still air box to settle for a few minutes.
Wipe down all the utensils you will be using for the inoculation with alcohol and place them inside the still air box.
I like to keep a small dish of alcohol inside the still air box as well to wipe down utensils if I feel they need it.
Inoculate Liquid Culture With Agar.
Wipe your hands with alcohol again.
Take your scalpel and move it outside your still air box, flame sterilize the end until it is glowing red then move it back inside the still air box.
Wipe the handle with alcohol again (if you have pre packed sterile blades you can just put a new blade on inside the still air box as it will be sterile).
Put everything inside, close the lids and allow the still air box to settle for a few minutes.
Remove the para-film tape from the outside of the agar plates.
Using the scalpel cut a piece of mycelium out from the agar then in one smooth motion move it over and drop it into the liquid culture jar.
Seal the lid of the liquid culture jar back up. Then move the jar into the incubation.
Incubate Liquid Culture.
Incubate your liquid culture at the appropriate temperature for your mushroom strain. For most strains this is around room temperature.
If, like me, you have a stirring pad and placed a stirring rod inside your liquid culture, then place your liquid culture jar onto the stirring pad once a day to break up the mycelium.
If you don’t have one of these you can give the liquid culture a shake or just let it form into one large mass in the middle.
I have details about how to break up the mass of mycelium on “how to use liquid culture” if you end up in this position.
The mycelium can take several days to a week to fully colonize from here.
It's important to monitor the progress of your liquid culture and look out for any signs of contamination.
Checkout how to make liquid culture for a picture on what contamination looks like in comparison to a good jar.
Once the liquid culture is fully colonized, it can be used to inoculate larger cultivation substrates such as grain spawn or bulk substrates.
Conclusion
Making liquid culture from agar is a simple and efficient process that allows for expansion of mycelium for mushroom cultivation.
By using a sterile technique one can easily create a liquid culture that can be used for multiple growing projects.
It is an important step in mycology to learn how to make your own liquid culture and can produce unlimited spawn for growing.
