|By Nan (Nanook) on Friday, October 26, 2001 - 03:18 am: The Nook|
Sterile Tissue Cloning Procedure
1. Create a paste of brown rice flour and water (or your choice of substrate), spread evenly in the bottom of a canning jar, seal, pressure cook at 15 psi for 30 minutes.
2. Pour about 1/3 cup of 3% h2o2 into a jar.
3. Obtain a clean coffee grinder*; fill with water to the brim of the metal cup, insert a fresh and cleaned mushroom stem (no cap) and proceed to grind for 10-15 seconds.
4. Use a clean spoon to transfer some of the resulting mixture into the h2o2 jar.
5. Let it fizzle for a couple minutes, then draw into a syringe.
6. Inoculate jar created in step#1, or simply inoculate pf-style jars.
Tips if Culture Contaminates
1. Extract a piece of healthy mycelium.
2. Cut mycelium into tiny pieces with razor blade.
3. Transfer pieces to a small pool of h2o2 and then suck into a syringe.
4. Inoculate another culture jar with it.
Stolen from Learner
* Notes from Nan: A Blender is much better than a coffee grinder.
Also Dirty Prints and other cultures can sometimes be cleaned by germinating a small quantity of the contaminated spores (or inoculating with some other contamed culture) in a clear liquid culture, then go fishing with a big needle. Gently suck up some clean mycelium into a sterile syringe pre-loaded with 5% solution of 3% H2O2 (one part H202 3% in 19 parts Sterile Water). Suck in a cc or two of air and shake the hell out of the syringe. Wipe the needle with alcohol, then let the syringe sit for 10 minutes with the needle up and covered (it will squirt if the needle is not up). Shoot clean Dextrose jars and some Agar Plates at the rate of 1/4 cc per culture.