How to Inoculate Mushroom Substrate Bags With A Liquid Culture Syringe Liquid cultures are used to start mushroom growth on almost any medium including fruiting bags, grain bags, plugs, and agar. They are the most convenient way to start culturing new species. The following instructions show grain bags being inoculated, but the same process can be used on any bag. Step 1 - Gather Supplies You will need a flow hood or glovebox, 70% alcohol, paper towel, nitrile gloves, a roll of scotch tape, your culture, and sterilized grain bags. Turn on the hood. Step 2 - Clean Hands Put on the gloves, then spray them with alcohol generously. rub them together well, be sure to get the top of your hands and between your fingers. Step 3 Clean Prep Pad or Towel Spray alcohol on a paper towel, be sure to cover it well, but it does not need to be dripping. Step 4 - Wipe Injection Point Wipe an area or spot on the substrate bag which you will be injecting through. Do not wipe or inject through the white breather filter on the bag. Step 5 - Shake Syringe to Mix Culture Keep the cap on. Shake your culture syringe hard for 30 seconds. This will help break up the mycelium and spread cells throughout the syringe. Step 6 - Open Needle Packaging Open the needle by peeling back the paper and plastic. Try not to touch what is inside. Remove the cap from the culture syringe and twist the needle tight into place. Step 7 - Remove Needle Cap Next, prepare for inoculation by pulling the clear cap off to expose the needle. Step 8 - Inject Substrate Bag Pierce the bag with the needle where you prepped it with alcohol, and inject 1ml of liquid culture into the bag, then withdraw the syringe. Step 9 - Take Clean Tape Take a 1-inch piece of tape. Be sure to remove the first couple inches of previously exposed tape when you first enter the hood or glovebox in order to eliminate any contaminates. Step 10 - Cover Injection Point Cover the injection point with the fresh tape, be sure it is sealed well. It is good measure to grab a second piece of tape and cross over the first. Step 11 - Mix Substrate Bag After the injection point is sealed, unroll and mix your substrate bag. Mixing the bag spreads the culture through it resulting in faster incubation times. Step 12 - Incubate Until Ready Set your inoculated bag in a dark place between 70F-90F (80F is best for most species). This process takes 12-30 days depending on the species.