Inoculating Liquid Culture with Agar

Cloning a wild harvested mushroom has a high risk of contamination. As it is possible to isolate mycellium away from contamination (sectoring) when using agar media if you begin the process using agar you can then be sure of having a pure culture to use as an inoculum for you liquid media. In essence it is a similar process to cloning a mushroom but you are extracting the mycellium from the surface of the agar.

Materials:

  • 1 airport jar with steriles liquid meida
  • 1 airport jar with fresh water
  • 1 syringe with needle (16-18 guage)
  • Tin foil
  • Alcohol spray (ISO Propynol diluted to 70%)
  • 1 fresh mushroom

 

Method:

  1. Sterilise the jars, syringe and needle (if re-using glass syringe and steel needle). Allow to cool.
  2. Aerate Liquid media jar (see Liquid Culture Growth and Development)
  3. Spray both silcone injection sites with alcohol.
  4. Using syringe draw up 5ml of water.
  5. Open the petri dish, working quickly and minising contact, inject 1ml of water onto the leading edge of the mycellium.
  6. Using the needle gently agitate the mycellium to suspend it in the water; suck up the water and mycellium intothe syringe.
  7. Insert the needle through the airport lid and inject in the jar.
  8. Label and date the jar to be stored in your incubation space

 

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