Try your best to disturb the bag as little as possible during this time period. You should see signs of growth 3-10 days after inoculation. It doesn't matter if it is dark or light. Step 5: Place your bag in a place that is subject to minimum disturbances and will maintain a temperature between 75 and 81☏. Step 4: Carefully inoculate the bag through the self-healing injection port with 1.5-2 ccs of spore or culture solution. If a lighter is used instead, wipe off any lighter soot that is left behind on the needle with a sterile alcohol swab immediately after flame sterilization. This step is best performed with the assistance of an alcohol burner because it will not leave any soot on your needle. After flaming the needle, press the plunger of the syringe down slightly and squirt a small amount of solution out of the needle to cool it down. Step 3: Flame the needle of your syringe for 15 seconds. Wipe down the black self-healing injection port with a sterile alcohol swab. Step 2: Put your gloves on if you have not already. You want to make sure to "inflate" the bag enough so the filter has some room to "breathe." If the filter is pressed up against the plastic of the bag, it will limit gas exchange during the colonization phase and could potentially stall the colonization process. Carefully pull the gussets on the bag apart all the way up to the top of the bag, where it is sealed. Step 1: The first step of inoculating a grain bag is to prepare it for injection with spores or mycelial culture. It is also recommended to work in a clean, small room with the door closed to prevent airflow. If you are going to work in open air, we suggest turning your AC/heater off an hour prior to inoculating your bags (this reduces the amount of airborne contaminants in the area). You should be doing the injection in a glovebox, or at the very least, a still-air environment.
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