• Boston Mycology Mushroom Grow Bag

• 3 mL Spore Syringe

• Isopropyl Alcohol Spray (70%)*

• Face Mask*

• Disposable Gloves*

• Knife/Scissors*

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*Materials not included in Mushroom Grow Bag Kit*


 

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1. Gather your materials in a clean, uncluttered workspace with no draftiness.

   1. You will want to complete this process in a room that does not have a draft or active airflow. A still-air environment reduces the chance of contamination from pathogens landing on your workspace while you complete the following steps. Turn off all fans and close any windows to allow the air in the room to sit undisturbed for a few minutes.

2. Wash your hands and don your mask and gloves. Spray gloves with alcohol and allow to dry.

   1. It is important to complete this step before continuing, as your hands pose the highest risk of contamination during this process. Any bacteria on your hands or the gloves can pose contamination risks, and thus, this risk must be mitigated before continuing.

3. Wipe down your workspace of any debris and then sanitize the space with isopropyl alcohol.

   1. You can apply alcohol directly to a paper towel and wipe the space down, but the most effective method is to use a spray bottle and then wipe the surface with a paper towel after. When applying the alcohol, it is important to note if your mixture is 70%, 90%, or even higher. If it is not a 70% mixture, you will want to dilute the mixture down until it is roughly 70% using H2O. Though this may seem counterintuitive, the higher water content in the alcohol causes it to evaporate slower, increasing surface contact time and more effectively killing bacteria and viruses.

4. Spray each individual item and place them onto the sanitized workspace.

   1. Like the previous step, it is important to sanitize all items used in the process to best reduce contamination risks. After you spray each item, you can place it into the workspace and allow it to sit and properly sanitize.

5. Take the syringe and inspect it. If it does not appear damaged or compromised, give it a firm shake to stir the liquid culture.

   1. Making sure your syringe is not damaged or compromised is important, as any cracks, large air pockets, or other irregularities can lead to contamination of the spores and thus a spoiled product.

6. Locate the injection port on the bottom of the bag. Remove the cap of the syringe and carefully puncture the center of the injection port and plunge the syringe in about ⅔ of its own length. Push the end of the plunger and inject all of the liquid culture into the bag.

   1. When completing this step, make sure you use a careful and steady hand, as a fast or forceful puncture could rip the bag and lead to contamination. For those of you feeling confident with your handiness, you can slowly move/swirl the syringe around as you inject it to more effectively spread the spores, but be careful as to not slide the syringe in or out any deeper while doing so.

7. Slowly remove the syringe and give the injection port a final spray or wipe with isopropyl alcohol.

   1. Congratulations! You’ve completed your first mushroom inoculation and are on the fast track to becoming a mycologist. Take a second to revel in your work and think about what will come next (it’s important for you AND your new mushroom pal).

8. Locate a dark, warm space for your bag and place it there.

   1. You will want a spot that has a consistent temperature within a suitable range for your particular species. Your spore syringe should have included a sheet with temperature ranges best suited for the mycelium as it colonizes the grain and then substrates. You want to ensure that there are no drastic temperature fluctuations, especially those above the suggested range. Temperatures above the ideal range increase contamination risk and put stress on the mycelium, meaning that it will have trouble inoculating into the grain and then colonizing the substrate.

9. After about a week, check on your grain. Carefully lift the bag and inspect for any signs of colonization. If there are still large portions of grain that have not been colonized, allow the bag to continue for another week before moving to the next step.

   1. Mycelium will be white and appear as fluffy patches. If the mycelium is stressed, it may secrete a beige/yellow/brown liquid as it continues to colonize the grain. A little bit of this is okay, as it is just the digestive enzymes of the mycelium, but if there is a larger portion of liquid than mycelium or you cannot see mycelium, you will need to evaluate your environmental conditions and likely find a new spot for your bag. If any other color is present, including green, red, black, yellow, or orange, this is likely contamination, and thus, the bag must be discarded.

10. Once the grain has been fully colonized, carefully break the grain up with your hands on the bottom of the bag.

    1. Be careful to not puncture the bag while doing so, as the grain can be firm and thus require a reasonable amount of force to break apart. The best way to do this is to squeeze the grain from the sides and squish it in on itself.

11. With the grain sufficiently broken up, hold the bag by the bottom and shake the contents. Shake until the contents are evenly mixed and then grab the top of the bag and shake the bag once more to settle all the contents back to the bottom of the bag.

    1. You will want to be thorough when mixing the substrate and grain, as even distribution of the mycelium ensures proper colonization and reduces contamination risk.

12. Once again, locate a dark, less warm space for your bag and place it there.

    1. For this colonization stage, you will want to find a space that is a few degrees cooler than the initial spot you found. It should be a darker space as well, but as your mycelium matures and prepares itself for fruiting, the cooler temperature is more conducive to their natural environment and thus will allow for better substrate colonization. Once again, this temperature range will be included on the sheet that came with your spore syringe. If you do not have a new spot, that is okay, but make sure to keep an eye on the mycelium to ensure it is colonizing properly and there is no contamination.

13. Continue to monitor the bag until it becomes fully colonized.

    1. This will be obvious, as the bag goes from brown to white once fully colonized with mycelium. Sometimes your mycelium will refuse to colonize a small portion of the substrate, usually on the bottom, and that is okay. If this is the case, monitor the patch for a week. If there are no changes over the course of a week, that means the bag is ready to fruit.

14. Once your bag has been fully colonized, locate a new spot with dappled sunlight and that is within the temperature range.

    1. Though we have emphasized the importance of locating a spot within the ideal temperature range, for no step is this more important than the fruiting stage. Temperature, humidity, and fresh air are all essential to fruiting body production, so if any of those environmental conditions are not ideal, the mycelium will not fruit.

15. Place your bag into the new location and, using a sanitized knife or scissors, cut a slit into the bag. Fold the top of the bag over on itself, squeeze excess air out, and secure it with a piece of tape.

    1. You need to ensure that its opening is both large enough to allow for airflow across the face of the mycelium while not too large to where it would dry out the mycelium. Typically, a square about 1 inch on all sides or an X about two inches across will be a sufficient opening.

16. Using a spray bottle, generously mist the opening of the bag twice daily until you notice pins beginning to form. After those begin to form, transition to lightly spraying the bag once a day.

    1. Pins typically take one to two weeks to form. After pinning has begun, the mushrooms will take about another week to reach full size. As they mature, you may notice color changes.

17. As your mushrooms grow, monitor them for signs of maturity that indicate they may be ready to harvest.

    1. Mushrooms reproduce via spores, so monitor any changes to them that may indicate they are preparing to drop spores. This can be obvious, like with Blue Oyster Mushrooms and the caps unfurling, but for others like Pink Oyster or Lion’s Mane, it is not as apparent. Visit the Boston Mycology website for photos of harvestable fruits and feel free to research even further using resources found online. No two mushrooms are the same, and thus, you must use your mycological instincts here. You've gotten this far, so be proud and confident in your ability and your work, as you can now call yourself a mycologist!

18. Often, mushroom fruiting bags can produce multiple flushes of mushrooms, so repeat Steps 14–17 for a new flush.

    1. During the next few flushes, you will notice the fruiting bodies will be smaller in size and sometimes produce fewer larger individual mushrooms instead of the large cluster like the first time. This is normal, so don’t worry about their appearance, as they will cook and taste all the same. If your bag appears to be stalling out but you have only completed a single flush, you can briefly soak the bag in water to rehydrate the substrate, but make sure to only do this for 30 minutes to an hour as to not drown the mycelium.