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How to Use a Colonized Agar Plate
What You Will Need to Get Started
To get started using your fully colonized agar plate you will need to gather up some basic supplies. By doing this and following sterile procedures it will help increase your chances of success. When dealing with a fully colonized agar plate it is essential that all work is done in front of a laminar flow hood or in a still air box. Not using either one of these tools will most certainly result in failure. Please see equipment list below.
- Fully colonized agar plate with your culture of choice.
- A still air box or laminar flow hood.
- Scissors or equivalent to open the bag.
- A way to sanitize the agar plate, rubbing alcohol or equivalent. Most sanitizers work.
- A scalpel. The longer the handle the better we have found in our experience.
- A propane torch or equivalent to sterilize the scalpel blade.
- A sterilized mushroom substrate jar or bag, blank master slant, liquid culture jar or blank agar plate to transfer the agar wedge to for inoculation.
- FIRE AND ALCOHOL DO NOT MIX! Use caution and be aware of your surroundings at all times when dealing with open flames and especially with flammable liquids out.
- Not doing so can result in property damage, injury or even death.
- Keep all flammable liquids and materials far away when you have an open flame.
Now that we have gathered up all of our supplies it is time to begin. In this example we will be inoculating hydrated and sterilized rye berries we prepared for this tutorial. For one quart of sterilized rye berries we recommend you add four to five - 1 inch colonized agar wedges.
Sanitize the work area and your substrate bag or jar.
Fully wipe down the substrate jar.
Dirt and bacteria can hide in small places.
Dirt and bacteria can hide in small places.
Fully wipe down the substrate bag or jar you plan to inoculate. Use rubbing alcohol or equivalent. Most any sanitizer will work.
Dirt and bacteria can hide in small places.
Dirt and bacteria can hide in small places.
Dirt and bacteria can hide in small places.
The more thorough you are in your sanitation techniques the better. Dirt, dust and bacteria are everywhere naturally, Keep it clean!
Preparing your Colonized Agar Plate
Cut Open the Vacuum Sealed Bag
Cut Open the Vacuum Sealed Bag
Cut Open the Vacuum Sealed Bag
Your colonized agar plate will arrive vacuum sealed. You will need to cut open the sealed bag to remove the agar plate.
Remove the Agar Plate From Bag
Cut Open the Vacuum Sealed Bag
Cut Open the Vacuum Sealed Bag
Having just cut open the bag, reach inside and remove the fully colonized agar plate from the vacuum-sealed bag.
Remove the parafilm and sterilize the scalpel.
Remove the parafilm around the agar plate.
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
Find the end of the parafilm on the agar plate. Gently pull and allow the agar plate to rotate. This will allow the parafilm to easily unravel.
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
While using extreme caution, now flame-sterilize the scalpel blade until it is red hot. This will ensure no contaminants are on the blade.
Get set up and cut the agar wedge.
Remove the substrate jar lid.
Carefully hold the plate and cut the wedge.
Carefully hold the plate and cut the wedge.
We placed our sterilized scalpel on a jar ring to cool. Remove the lid and prepare to transfer the agar wedge to the mushroom substrate.
Carefully hold the plate and cut the wedge.
Carefully hold the plate and cut the wedge.
Carefully hold the plate and cut the wedge.
Carefully hold the agar plate with one hand and cut a small 1-inch wedge. We prefer to use a triangle cut, but do what's comfortable.
Add the colonized agar wedge to the substrate.
Place the agar wedge on top of the substrate.
Place the agar wedge on top of the substrate.
Place the agar wedge on top of the substrate.
Gently move the colonized agar wedge over to your substrate jar. Make sure to keep the wedge in the sterile air flow during the transfer.
Drop in the Agar Wedge
Place the agar wedge on top of the substrate.
Place the agar wedge on top of the substrate.
Drop the colonized agar wedge into the substrate jar. You may need to convince it to fall by scraping it on the inside of the jar rim lightly.
Lets Finish Things Up
Replace the Lids and Put Your Tools Away
If you are adding colonized agar wedges to one quart of substrate, like in this example, we recommend you use 4-5 agar wedges per jar. Shake the jar a few times to distribute the wedges. If they get stuck on the lid or inside of the glass, it doesn't matter, just leave them. Do not open the jar again until it has fully colonized. In most cases agar wedges produce some growth with in 3-4 days and at least 2 weeks until fully colonized depending on variables such as mushroom type, temperatures, how many wedges used, and the type of substrate used. Record your data!
Replace the lid to the sterilized mushroom substrate jar and the colonized agar plate. Wrap the sides of the agar plate with para film or saran wrap. An additional measure against drying out, we recommend you then place your colonized agar plate inside of a zip lock type sandwich bag.
Clean up your work area. Remember there is a open flame source, flammable liquid and a open surgical blade laying around. Take the time to clean up and put everything away. The last thing you want is a pet or loved one to get hurt by these tools being left out and unattended.
How you store your colonized agar plate for long term depends on the type of mushroom it is. Some mushrooms can tolerate cold temperatures where as cold will kill other mushrooms. Please fully research the mushroom you are working with to see if you should use room temperatures or the refrigerator for storage. Most tropical mushrooms don't like the cold.