Place all of the following items into the Glove Box.
A nice size mushroom. Remember, you will be tearing the mushroom stem down the middle.
New X-Acto knife (hobby razor)
Sterile Agar petri dish(s)
91% Alcohol in a small cup
Small alcohol soaked paper towel
Sandwich size Ziploc (one for each petri dish)
Cloning is so easy to do you will not believe it!
Everything that I am going to show you should be done within a Glovebox. There is no way to do this without a complete sterile environment.
And away we go!
Plug in the Glove Box that has been prepped for usage (if applicable)
Place the prepared Agar/petri dishes in the Glove Box. Also place your mushroom, alcohol soaked paper towel…etc into the Glove Box. After placing everything into the Glove Box go ahead and wash your hands with some antibacterial soap.
Dip your razor into the alcohol and flame it with your lighter. Place it on the alcohol towel in order to keep it clean.
Tear your mushroom down the stem and carefully pick a small piece of flesh from the inside of the stem with your razor and place it in the center of the petri dish and quickly replace the lid. Dip your razor into the alcohol and flame it again. This will eliminate the chance of “cross-over” contamination. Repeat the process for a total of three petri dishes in order to guarantee yourself at least one clean petri dish. If you like you can place a piece of masking tape over opposite ends of the petri dish before placing it into the baggie. It is not necessary, but it will hold the dish together to reinsure that it does not accidentally open and become contaminated.
Place each petri dish into separate baggies or all three into one.
Place the petri dishes into a warm place, your underwear drawer is good, and check on it once a day. You will notice that the mycelium becomes fuzzy and begin to spread over the surface of the dish. You should be seeing growth in about 2-3 days.
If it is contaminated (and you will know due to the green or black fungus) not all will be lost. Just repeat the same steps, except, you will be transferring non-contaminated mycelium to a new petri dish. Do this until you have a non-contaminated dish.
When the mycelium reaches the edge it is time to use or store the mycelium in the refrigerator (upside down) until you are ready to use it.