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old hand Reged: 11/21/04 Posts: 9056 Loc: Somewhere Else |
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:LIQUID CULTURE - THE EASY WAY: (how to turn 2ml/cc of spore solution into 20 ??..ten ml/cc syringes - right - cheap & easy. Assuming you have an empty syringe & another with spore solution. Go to Kmart, Target or any large discount Mart & buy a 3 pack of EvenFlow plastic baby bottles. Also buy a small bottle of Karo corn syrup, a tube of silicone sealant & a few glass marbles. All the baby bottle parts (bottle - screw cap - nipple) will withstand sterilization in a pressure cooker. Remove the nipple, apply a small amount (1/8th to ? inch) of silicone sealant to the inside (interior) of the nipple, to serve as a self sealing syringe needle injection point. Allow the sealant to cure overnight Note: the baby bottle has milliliter (ml) graduation marks up to 240 ml (8 fluid ounces). Add 200 ml of water to the baby bottle (use marks on it - as a measuring point). Take your empty syringe and aspirate 6 (six) ml of karo syrup into it. Then inject that into the open baby bottle, resulting in an approximate 3% solution. Next, add a glass marble to the baby bottle, to serve as an agitator devise - later. Insert the nipple into the baby bottle (facing down - inside the baby bottle). Attach the screw ring lid to hold the nipple in place over the mouth of the baby bottle. :IMPORTANT: Attach the screw ring loose, to allow steam to escape & pressure equalization during pressure cooking. Pressure cook the baby bottle (or several) 20 minutes @ 15 psi. :IMPORTANT: Allow the pressure cooker to cool to room temperature, otherwise a vacuum can form inside the bottle, if the screw ring is tightened while the bottle is hot.. Open the cooled pressure cooker & immediately tighten the screw rings on the bottle (or bottles). You now have a sterilized nutrient solution, in which to inject 2 or 3 ml/cc of spore solution into. Once you inject a spore solution into it, simply incubate it around 78 to 82F in a clean environment. You should be able to see tiny strands of mycelium (from germinating spores) in the solution within several days. Be patient, as some spores take longer to germinate - than others. Once you have visible mycelium in the culture, gently agitate in on a daily basis. Once the mycelium in the culture is fairly thick, it is ready to be aspirated into sterilized syringes, and used an inoculate. ~~~~~~~ FOR THOSE WITHOUT A PRESSURE COOKER: NOTE: Not as fool/proof - fail/safe as pressure cooking sterilization. Cut a piece of tyvek in a circle. Use a rubber band to hold it in place over the screw cap of the baby bottle. Insert the baby bottle into your microwave (be sure the lid is somewhat LOOSE). Then, microwave heat it until the solution in it - comes to a boil. Then - stop, wait a moment & do the same thing again. Repeat, several times. Remove the baby bottle (gently - as the lid is still loose) & allow to cool to room temperature. Then, tighten lid. ~~~~~~~~~~~~~~ Or, if you want to do gallons, use jars - with filter lids. --------------------
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