501 lines
20 KiB
Markdown
501 lines
20 KiB
Markdown
# HomeMyco
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A [homebrewer](https://en.wikipedia.org/wiki/Homebrewing)'s take on practical home cultivation of [mycelium](https://en.wikipedia.org/wiki/Mycelium) and their [mushrooms](https://en.wikipedia.org/wiki/Mushroom).
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# Always a Risk of Contamination
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Contamination occurs when another microorganism takes advantage of the gratuity you provide for your mycelium.
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There is always a risk of bacterial and mold contamination when it comes to mycelium.
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While many guides do everything they can to reduce this risk, my approach is to make the methods quick, cheap, and simple.
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Contaminated spawn is easily discarded and replaced.
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# Do It Thrice
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As a result of the above hubris we triple our efforts.
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- When making [colonization jars](#grain-spawn) make at least two for each sample we wish to [colonize](#colonizing).
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- When making [agar slants](#agar-slants) or [liquid cultures](#liquid-cultures) make at least two for each sample.
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- Make and save at least one of the above as a control for testing your sanitation.
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It is OK.
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Seed spawn, agar, and sugar water are inexpensive.
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Saving a control spawn medium, both [grain spawns](#grain-spawn) and [culture mediums](#culture-mediums), will help you test for contamination introduced through your technique.
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If all fares well you can always end up using it.
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# Finding Samples
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- [ ] mycelium sample
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At the beginning of each section is a checklist of what to buy.
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At the end of the document there will be a combined checklist.
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Newbies should start with store bought mushrooms.
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This will allow you to walk through the methods without fear of wasting a sample.
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Getting other samples should not be difficult but it is quite frustrating to discover all if the sanitary steps were made in vain when a contaminated sample was the source.
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Of course, there is always the risk of a contaminated store bought sample.
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Online shopping can get you all sorts of sample types ranging from spawn grains and liquid culture to spores and dried mushrooms.
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When paying good money for a sample try to find liquid culture as they have a pretty good colonization rate.
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Eventually you will want to make your own cultures from the spores of your harvested mushrooms.
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# Sanitize Your Workspace
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- [ ] empty spray bottles, chemical resistant
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- [ ] star san, sanitizer
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Prior to working with a mycelium sample or spawn medium you should clean your work area.
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All surfaces and tools should be wiped clean then sprayed with a sanitizing solution at least 2 minute prior to working.
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Purchase a spray bottle [resistant to chemical reactions](https://en.wikipedia.org/wiki/Chemical_resistance) for this purpose.
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These can be found at most hardware stores.
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While you are at it you should buy a second spray bottle for misting water.
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If you are working near your living space a food safe sanitizer like [Star San](https://fivestarchemicals.com/star-san-sanitizer-4-oz) avoids stinking up the place or contaminating food.
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It is very concentrate so just a couple drops into a spray bottle full of water be enough.
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You will know if the solution is diluted enough when it sprays easily and creates small bubbles on the sprayed surface.
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It is important to note that sanitization is not sterile.
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Star San does not kill all bacteria or mold.
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There is always a risk of contamination.
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Your face, hands, and tools are the major source of contamination.
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Tools can be sanitized in a pressure cooker or [autoclave](#autoclave) which is discussed later.
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Your face, hand, and the other hand are a little more difficult to contain.
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While gloves and a face mask are often recommended I find they make me more clumsy to the point of negating any benefit they might provide.
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Face masks just do not work unless they provide respiration filtering.
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Gloves only work if they begin sterile and remain sterile.
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Scrub your hands with warm water and soap to remove anything that might fall off into the spawn medium.
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Avoid touching your spawn medium and the part of your tools that interact with the spawn medium.
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Wash and dry your face with warm water and soap to remove anything that might fall off into the spawn medium.
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Combing your hair and wearing a hat helps to reduce the amount of lose hair that might fall into the spawn medium.
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Slow or hold your breath when inoculating spawn medium to avoid shaking.
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Remember that mistakes happen and the look pretty cool when they do.
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# Three Stages
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There are three general stages to cultivating mycelium.
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1. [Culturing](#culturing), [wiki](https://en.wikipedia.org/wiki/Microbiological_culture)
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- Using simple sugar and agar as a surface to proof sample mycelium for viability and contamination.
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2. [Colonization](#colonization), [wiki](https://en.wikipedia.org/wiki/Colony_(biology)#Microbial_colonies)
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- Using grain as food source to promote mycelium growth throughout.
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3. [Fruiting](#fruiting), [wiki](https://en.wikipedia.org/wiki/Sporocarp_(fungi))
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- Apply the ideal conditions to fully colonized grain spawn to promote the growth of mushrooms.
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# Culturing
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Any sample of mycelium is potentially contaminated.
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Aside from using a microscope and checking the entire sample, using a small culture will allow you to grow a test patch of your sample without risking a whole jar of [grain spawn](#grain-spawn).
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The two [culture mediums](#culture-mediums) preferred are [agar slants](#agar-slants) and [liquid cultures](#liquid-cultures) together.
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While petri dishes are part of the usual trappings of cultivating the growth of microorganisms they are fairly high maintenance and better suited for a laboratory environment.
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Agar slants provide the same functionality without the need to seal and re-seal with parafilm.
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[Liquid cultures](#liquid-cultures) provide a similar micro climate as [agar slants](#agar-slants) with the added benefit of being a much easier for inoculating [grain spawn](#grain-spawn).
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The disadvantage of liquid culture is being unable to isolate mycelium from contaminations.
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You should use [agar slants](#agar-slants) to isolate mycelium from contamination then sample the mycelium from the agar to liquid culture.
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# Culture Mediums
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- [ ] food scale, grams
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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] distilled water
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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave)
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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## Agar Slants
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- [ ] malt extract agar
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Agar agar is a plant gelatin used to provide a physical surface that mycelium can colonize.
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This is mixed with a simple sugar like malt extract to provide mycelium calories for reproduction.
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Malt Extract Agar (MEA) is available in a premix fairly cheap online.
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Mixing 2:3 grams ratio simple sugar to Agar powder is good for saving money in bulk.
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Scale the following recipe as needed to fill 1/3 of each test tube or 10 ml:
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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1. Fill a 16 oz mason jar with 100 ml water on a food scale.
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- Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure).
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2. Microwave with a safe cover for 1 minute or until 50 celsius or higher without boiling.
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3. Add 5 grams of MEA to warm water and stir for 1 minute with a sanitary utensil.
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4. While the MEA solution is still warm, fill a sanitary syringe with the solution.
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5. Fill a sanitary test tube one third (1/3) capacity and replace the screw cap.
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6. Place test tubes in an empty mason jar and [autoclave](#autoclave) for 1 hour.
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7. Carefully remove each test tube and place on a flat surface with the screw cap slightly elevated on a dish towel.
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- Once cooled the agar should solidify into a slanted surface for inoculation.
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8. Store in a dark cool place until inoculation.
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Do not freeze.
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## Liquid Cultures
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- [ ] light corn syrup
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Mixing liquid culture is similar to [agar slants](#agar-slants) without needing to cool in any particular position.
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A simple sugar like light corn syrup is dissolved in distilled water to provide mycelium calories for reproduction.
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Scale the following recipe as needed to fill 2/3 of each test tube or 20 ml:
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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1. Fill a 16 oz mason jar with 100 ml water on a food scale.
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- Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure).
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2. Microwave with a safe cover for 1 minute or until 50 celsius or higher without boiling.
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3. Add 5 grams of light corn syrup to warm water and stir for 1 minute with a sanitary utensil.
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4. Fill a sanitary syringe with the solution.
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5. Fill a sanitary test tube one third (2/3) capacity and replace the screw cap.
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6. Place test tubes in an empty mason jar and [autoclave](#autoclave) for 1 hour.
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7. Store in a dark cool place until inoculation.
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8. Once inoculated, shake the test tubes daily to oxygenate the water and break up the mycelium.
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## Autoclave
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[Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for a pressurized steam chamber for sanitization and sterilization.
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**All autoclave duration measurements use electric pressure cookers for convenience.**
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If you have a stove top pressure cooker or industrial autoclave that provide a continuous 100 kPa (15 psi), reduce the time by 1/3.
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Autoclavable tools are safe to bring to 250 celsius for extended durations without damage.
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Use the following durations as they make sense to sanitize and pasteurize.
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- 2 hours
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- Spawn grain should be autoclave for 2 hours prior to inoculation.
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Bulk grains are particularly susceptible to contamination during the commercial farming process.
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- 1 hour
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- Cultures do not need to autoclave for as long as spawn grain since there is less volume that needs to get to temperature.
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- Add your autoclavable tools if there is space.
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If you do not plan to use them soon after autoclave, soak in Star San solution and store covered.
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- 10 minutes
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- Quickly bring test tubes and tools to temperature for cleaning out old agar or liquid culture.
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0. Add the steam rack to the inner pot of the electric pressure cooker.
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1. Add 200 ml of water.
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2. Insert items to be autoclave.
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3. Press Steam option and set the appropriate time.
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4. Turn vent valve to close.
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5. Quick release once time has completed if needed, working with agar warm is easier.
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# Culture Inoculation
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Once you autoclave your [culture mediums](#culture-mediums) and they have cooled to room temperature it is ready for mycelium.
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Depending on the source of the sample there are two ways to inoculate, [cloning](#cloning) from a mushroom or using [spore samples](spore-samples).
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Newbies should start with store bought mushrooms.
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This will allow you to walk through the methods without fear of wasting a spore sample.
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## Cloning
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- [ ] paring knife or scalpel
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Taking a mycelium samples from the stem of a mushroom is called cloning.
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The collection of a sample for cloning is relatively trivial using a syringe with a hollow needle.
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [culture medium](#culture-mediums).
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1. With a sanitary knife, cut the stem off the mushroom then half down the length.
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2. Using the hollow needle tip of the syringe, plunge along the length of the stem collecting a sample of the core of the stem.
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3. Gently use air pressure from the syringe will push the sample into an open [culture medium](#culture-mediums).
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4. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature.
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## Spore Samples
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- [ ] inoculation loop
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- [ ] flame or lighter
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[Spore](https://en.wikipedia.org/wiki/Fungus#Spore_dispersal) samples are created by placing a mushroom cap on a flat surface and collecting the spores that are released.
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These are called spore prints.
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Spores can further be added to a syringe of sterile water for ease in shipping.
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In either case, an inoculation loop is used to apply the spore samples to a [culture medium](#culture-mediums).
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### Spore Print
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [culture medium](#culture-mediums).
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1. Using a lighter, heat the loop element of your inoculation loop until it glows red. Allow the loop to cool.
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2. Scraped spores off a print delivery medium with a sterile sharp.
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3. Collect spores on the loop.
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4. Insert loop into an open [culture medium](#culture-mediums) and gently apply spores.
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5. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature.
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### Spore Syringe
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [culture medium](#culture-mediums).
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1. Using a lighter, heat the loop element of your inoculation loop until it glows red. Allow the loop to cool.
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2. Place a single drop of water from a spore syringe on the loop.
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3. Insert loop into an open [culture medium](#culture-mediums) and gently apply spores.
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4. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature.
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## Wood Plugs
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- [ ] tweezers
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Mycelium samples are often sold as colonized grain or wood plugs.
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Usually these can be used to inoculate [grain spawn](#grain-spawn) directly but we can also make cultures to store these samples long-term.
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With sanitary tweezers place a sample in an [agar slant](#agar-slants), [liquid culture](#liquid-cultures), or grain spawn.
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# Reading an Agar Slant
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[Agar slants](#agar-slants) should show growth within the first week and should be monitored for contamination weekly.
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Mycelium are snowy white with grey and blue tints as normal.
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Any other colors and you are dealing with contamination.
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Contamination is not the end of the world.
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Mycelium can be isolated from a contaminated sample.
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If enough mycelium growth has occurred that a sample can be collected without touching the contamination then it can be transferred to a fresh [culture medium](#culture-mediums).
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [culture medium](#culture-mediums).
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1. Using a lighter, heat the loop element of your inoculation loop until it glows red. Allow the loop to cool.
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2. Gently scrape some of the mycelium onto the loop.
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3. Insert loop into an open [culture medium](#culture-mediums) and gently apply spores.
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4. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature.
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# Colonization
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Once the mycelium has been successfully isolated and grown in a liquid culture the mycelium wants something a bit more nutritious than sugar water.
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Most species have special requirements that encourage the growth of mushrooms.
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Wood-loving species like oyster and shiitake thrive in a mixture of wood chips.
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This guide does not address these special requirements.
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I leave it up to you to research your mycelium species.
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[Grain spawn](#grain-spawn) is a common source of nutrients and the most accessible source of grain spawn is wild bird seed.
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# Grain Spawn
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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] wild bird seed with millet
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- [ ] colander
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- [ ] ladle
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- [ ] boiling pot
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- [ ] aluminum foil
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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Wild bird seed is rather dusty and full of bacteria and molds.
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Because the seeds are dry the bacteria has likely [endosporulated](https://en.wikipedia.org/wiki/Endospore) making it resistant to heat.
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Soaking and boiling the bird seed will clean it up a bit and encourage the endospores to reproduce, leaving them weaker to autoclave.
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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1. Fill 16 oz wide mouth mason jars with 1 cup of bird seed per final jar of grain spawn.
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2. Cover bird seed with tap water, lid jar, and soak for 12 to 24 hours.
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3. Strain the bird seed with a colander and rinse with water.
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- Pick out any unwanted bits like twigs or mush.
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4. Bring a pot of water to a rolling boil.
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5. With a ladle, carefully add one scoop at a time to the boiling water.
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6. Once all the bird seed is added let the water return to a boil.
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7. Strain again with a colander for 1 hour to dry, stirring occasionally.
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8. Split evenly to each 16 oz wide mouth mason jar, about 1 cup.
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9. Cover un-lid jars with aluminum foil and [autoclave](#autoclave) for 2 hours.
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10. Carefully screw lid over the foil and jar and allow to cool to room temperature.
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11. Store in a dark cool place until inoculation.
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# Spawn Inoculation
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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each jar of [grain spawn](#grain-spawn) and [liquid culture](#liquid-cultures).
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1. With a sanitary syringe extract 1 ml of [liquid culture](#liquid-cultures) and replace the screw top.
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2. Inject the liquid culture into the jar of [grain spawn](#grain-spawn) by piercing the aluminum foil straight down along the side of the jar.
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3. Replace the mason jar lid by flipping it so the seal is facing up and lightly screwing on the rim.
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4. Store in ambient room light at room temperature.
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# Reading a Spawn Jar
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Similar to [Reading an Agar Slant](#reading-an-agar-slant), [grain spawn](#grain-spawn) should show growth within the first week and should be monitored for contamination weekly until it is fully colonized, 2-3 weeks.
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Mycelium are snowy white with grey and blue tints as normal.
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Any other colors and you are dealing with contamination.
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Minor contamination is not the end of the world, many colonies can recover.
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If the jar is fully colonized following a contamination and doesn't smell like rotten apples it can still produce mushrooms.
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# Fruiting
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If you've ever been walking in the woods after a light rain you've probably seen mushrooms peaking up.
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This is because mycelium sense that water is evaporating and wish to take advantage of the updraft to spread their spores.
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In our fruiting containers we want to mimic that as much as possible with a spray bottle and a light breeze.
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This can be accomplished with an open and fully colonized [grain spawn](#grain-spawn) jar and a daily spray bottle misting.
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However, if there is more surface area for the mushroom pins to sprout from there is a higher chance of successful fruiting.
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# Fruiting Chamber
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- [ ] medium plastic tubs with lids
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- [ ] coconut coir
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- [ ] personal fan
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Coconut husk (coir) as casing layer and flooring in our fruiting chamber to hold moisture better than the mycelium cake we created in the [grain spawn](#grain-spawn) jar.
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0. Invert an open and fully colonized [grain spawn](#grain-spawn) jar and firmly but gently tap cake out.
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- If the cake breaks up that is OK, place the pieces back together if possible.
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1. Soak the mycelium cake in water for 12 to 24 hours.
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2. Soak coconut coir brick following the packaging for the amount of water, about 15 minutes.
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3. Gently roll the mycelium cake in the hydrated coconut coir to form a light casing.
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4. Layer the bottom of the tub with an inch or more of coconut coir.
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5. Place the mycelium cake on the layer of coconut coir away from the sides.
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## Daily
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The goal is to maintain a 90% humidity within the fruiting chamber.
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0. With a water spray bottle gently mist the coconut coir and mycelium cake until the coir is moist but not soaked.
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1. Place the medium plastic tub lid upside down, slightly ajar, and rest for 1 hour.
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2. Point a personal fan nearby to blow towards the ceiling above the tub for 1 hour.
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3. Replace the lid upside down and slightly ajar.
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4. Harvest mushrooms before they dump their spores.
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# Checklist
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- [ ] mycelium sample
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- [ ] empty spray bottles, chemical resistant
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- [ ] star san, sanitizer
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- [ ] food scale, grams
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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] distilled water
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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave)
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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- [ ] malt extract agar
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- [ ] light corn syrup
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- [ ] paring knife or scalpel
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- [ ] inoculation loop
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- [ ] flame or lighter
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- [ ] tweezers
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- [ ] wild bird seed with millet
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- [ ] colander
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- [ ] ladle
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- [ ] boiling pot
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- [ ] aluminum foil
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- [ ] medium plastic tubs with lids
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- [ ] coconut coir
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- [ ] personal fan
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