Clean Spore Print
91% Isopropyl Alcohol
New Zipper lock Bag (Quart sized or larger)
Shot Glass or alcohol lamp
Match or lighter
The following procedure is an inexpensive method for making clean spore syringes without the use of a glove box. We used this technique for shiitake, but it can be applied to other mushrooms as well. Here is an overview of the process: You will be sterilizing water in a syringe, getting your print inside a clean bag, then injecting the sterile water from the syringe right into the bag and sucking it back out with the spores.
You will need an empty sterilized syringe to start off with. Fill your empty syringe with water and pressure cook it at 15 psi for 20 minutes. This will ensure that the water in the syringe is sterile. Water may be replaced with your choice of liquid growing medium if you would like the spores to germinate right in the syringe. You should be extra certain of the sterility of your print if this is the case. When sterilizing your syringes in the pressure cooker, make sure to keep them with the tip facing up or you will lose all of your liquid during the process. The caps should be placed loosely on top of the needle points so that you do not poke yourself when trying to grab them. You can use a quart size Chinese soup container to hold your syringes because it can withstand the temperature. Be sure to allow your syringe to cool fully before use.
Now that your syringe water is at a safe temperature for your spores, go ahead and soak your paper towel with rubbing alcohol. If you cannot find 91%, 70% will work. But try to use the highest possible concentration. Use the alcohol to sterilize the outside of your spore print. Keep it in between the folded halves of your alcohol-soaked paper towel. Then go on to sterilize the opening of the zipper lock bag. Most bags will do, but you want to try to get one that is not too thick, and you will see why later.
Once the outside of the bag is sterilized, you have to get your print inside without letting any air in. This involves closing all the doors and windows in your house and reducing body movement. You may want to spray the air with air sanitizer if you have it. The key is to only open up one side of the bag enough to get the print in. Before acting, prepare the print in the paper towel so that it will face the entrance of the bag properly and be able to drop right in. You want to be able to just open up the two halves of the paper towel and let it drop in. Once the print is in, close the bag up quickly and just give another quick wipe along the entrance with your paper towel.
Now is the part where a thinner bag with help you. You have to unfold the print while it is inside the bag. The ease of this part of the process will depend on your personal skill and how the print was prepared. Eventually, you will become good at it. The key is to be gentle and not use your fingernails too roughly so that they penetrate the bag. You will want the print situated at the bottom corner of the bag when it is finally opened.
Now that the print is open, you will need to sterilize your needle. Fill up your alcohol lamp or shot glass with isopropyl alcohol and light the top so you have a flame. If using a shot glass, you will have a better flame if it is filled to the very top. The flame will also give you some idea of how still the air is in your room. (You can utilize this earlier in the process when you are inserting your print into the bag as a gauge of your air movement.)
Next, you want to sterilize the tip of the needle so that the entire thing burns red hot. If you get one area to turn red, come back to it every second or two while you’re working on another area of the needle so that it stays hot. Don’t go too close to the plastic either, or it will melt. You may hear popping if there is any water inside the tip. When your whole needle is sterilized, cool it down by folding it in an alcohol soaked paper towel.
Hold your sterilized syringe in the towel and bring it to the ziplock bag. While it is safe inside the alcohol-soaked paper towel, use another part of the alcohol-soaked towel to wipe the part of the bag where you will be inserting your needle. (By the way, you may need to add alcohol to the paper towel as needed during the process).
From this point on, you want to act very slow and think out each movement before you act. Stick your entire needle into the bag through the spot you just sterilized with the alcohol-soaked paper towel. Do not worry about the direction the needle is facing. Instead, be sure that it does not go through the bag at any other point or it will let in unwanted air. If this happens, you may still succeed, but it is undesirable. You want the entire needle in the bag so that none of it is left in the open air to pick up contaminates. Once the needle is in, be sure to keep your alcohol-soaked paper towel over the entry hole so that no new air can get in. At this point you want to squirt out the contents of the syringe into the bag. You may squirt right on the spore print to remove the spores more easily, but that is not needed. The beauty of this technique is that you can rub the spores off with your finger right from the outside of the bag. Once the water is saturated with spores, slowly adjust your needle point to face the corner of the bag. Allow the water to collect in the corner and draw the water back into the syringe.
With this technique, you can usually get all of your spores into a single syringe. If you would like to do additional syringes on the same print, just be sure to keep the hole covered with your alcohol-soaked paper towel while you sterilize the next syringe.
That’s all there is to it…
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