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Northern spore9/23/2023 ![]() Step 1) To make slants, use a pipette to transfer 20 millimeters of PDA media into each glass tube. Pressure cookers vary, so make sure you follow the instructions to ensure you’re properly cooking your media. After, seal it again and let it go for another 18 minutes. Once the timer goes off, open the pressure cooker for a few minutes to air out. Step 4) Set a timer for 18 minutes (at this point you’ll see steam rising out). Close the pressure cooker properly by making sure each screw is balanced. Include any autoclavable bags you’ll be using to store the plates once they've been poured. After, take your solution, double check that it’s loosely lidded, and place it inside. We used an All American pressure cooker, but you can use any pressure cooker you may have at home for this step but ensure you follow the instructions for the particular model. Step 3) Once your solution is ready, prepare the pressure cooker. An optional step is to use a biological indicator to confirm your media is properly sterilized. Wait for the mixture to fully combine in solution and heat to temp. Step 2) While the water is heating up, measure out your dry ingredients and add them to the individual appropriate flask, Pyrex bottle, or jar following the recipes above. This allows any pressure or condensation to escape. Important: Seal your container but do not over tighten. Step 1) Fill your flask, Pyrex bottle, or mason jar with the appropriate amount of water and place on a hotplate. Liquid culture tends to colonize whatever substrate you’re trying to grow on a little bit faster than when you work with a plate. Because the mycelium is in a nutrient broth, it can contain lots of small hyphal fragments which can each serve as inoculation points when you use the liquid culture to expand onto other substrates. Liquid culture is less susceptible to airborne contaminants, as it is housed in a sealed container such as a syringe, and ideal for working with fungi at home or a slightly less sterile environment. Liquid culture is often how the public first encounters mushroom cultivation. Often, you’ll store your culture on slants and transfer a small portion of culture from the slant to a culture plate to begin work on it. The narrow opening of a test tube is a little harder to work with but less apt to capture contamination and allows you to culture a fungus on a small amount of solid media, place it in the fridge, and store it more or less indefinitely until you need it. Slants, on the other hand, are ideal for long-term culture storage. Plates contain solid media culture consisting of natural sugars and agar, a jelly-like substance derived from red algae that gives the mycelium an anchor while we do our work. Plates are useful for cloning wild mushroom strains, isolating specimens away from contaminants, and conducting mating trials between different strains. They allow you to observe growth patterns, speed of colonization, and give you the opportunity to separate different mycelium strains taken from the spores of a single mushroom. Plates are the work surface for working with fungi.
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