AGAR MEDIUM STERILIZATION AND POURINGI use my home made malts for making agar. After grinding one or so tablespoons of roasted grain in a coffie grinder I add it to a cup of water and bring it to boiling temperature, covered on the stove. About five minutes simmering should do. I let it settle and pour off the tea into a decanter with three tablespoons of agar. Fifteen psi for twenty minutes is plenty of time for sterilization. Let cool until decanter fask is cool enough to hold and place it directly into sterile space. Still chamber or flow hood. I use spray bottle of bleach to periodically spray inside my still chamber. Removing petri top with one hand and pouring with he other, fill each about a quarter inch with liquid. If the liquid is too hot condensation may appear on the tops. No big deal. Less time with the petri open means less exposure, and less chance of contamination.
Agar medium preparation Making malt "tea"Adding agarSterilisation Finished productGERMINATING SPORESStarting with a clean spore print is essential to this process. In sterile space prepare scalpel or inoculation loop, agar plates, spore print, and lighter.
Wearing gloves and using proper procedure to avoid vectors of contamination, sterilize scalpel with lighter (still box) or tool clave (recommended for flow hood). Scrape a a section of spore print and transfer the hundreds of thousands of spores onto the agar medium surface. ( Only about five percent may germinate) I like to draw a single line down the center of the dish. Make a smiley face, that's fun. Again, the longer the dish is open the greater the chance of contamination. So be swift. Once inoculated, dishes must be sealed with parafilm and labeled correctly. They may then be removed from sterile space.
CLONINGTaking a clone from a mushroom is a good way to ensure a strain is viable. Clone is taken from a fruiting body as it is the cleanest and newest growth. In sterile space, and sterile scalpel simply remove a interior wedge and place,it on agar. Some varieties are easier to propagate than others. Oysters are especially vigorous and easy to propagate.
Several days later mycelium begins to grow from the edges of the wedge.