What we mean when we refer to ‘cloning’ is not some complicated DNA procedure, but simply scraping tissue from a mushroom under sterile conditions, and then placing it onto sterile media to grow. Mushrooms are composed of mycelium, thus when we transfer a small piece of mushroom tissue to sterile media, we’re transferring live cells that will continue to divide, giving us in most instances, a perfect genetic copy of the original.
Firstly you will need to work under a still air box or in front of a Laminar Flow hood
- Pre made MEA plates
- Gloves, mask, hairnet etc
- Sanitise your work area with alcohol including your gloves, scalpel.
- Take your sample mushroom that you would like to clone, if the sample is big enough I like to wipe down the outside with alcohol.
- Tear your sample mushroom in half from the stem up (sometimes i use my scalpel to cut in a few mm so you can tear it more easily). Do not cut all the way through with the scalpel as you could transfer contamination from the outside into the area you will transfer from.
- Flame your scalpel blade and let cool.
- Choose a meaty part of the mushroom and cut out a small piece and transfer to your agar plate, this only needs to be a tiny amount of tissue (be careful if your mushroom has spores you want the flesh, not to transfer spores).
- You may want to transfer multiple pieces from different areas to ensure success, flame your scalpel between each transfer.
- Wrap your agar plate.
After a few days you should notice mycelium growing out from you mushroom tissue. After a week or two you should have decent growth. You may have picked up some contamination during this process (especially if you have tried to clone wild mushrooms) and doing multiple transfers is advisable. You should choose the healthiest, fastest growing mycelium and transfer each to multiple plates, eventually after around 3 transfers you should have clean mycelium that can be used to make grain spawn, liquid culture or for long term storage.
What Mushroom to Clone ?
If for example you have grown from a spore print you would have mushrooms growing with multiple different genetics, you would want to choose the mushroom with the most preferable traits to clone. When cloning, select a young, rapidly growing fruit for best results. You want a fruit that is rapidly dividing cells, so it will take off quickly on agar. I prefer to clone from clusters, because they’ll tend to produce clusters on future flushes.
Similarly with wild mushrooms try choose a healthy still growing mushroom with rapidly dividing cells, try clone as soon as you can after picking.
Older sporulating mushrooms should be avoided if possible, firstly because the cells stop dividing at this stage and secondly you don’t want to introduce the possibility of transferring spores with the tissue to you agar plate.