452 lines
19 KiB
Markdown
452 lines
19 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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You don't have to follow every step in this guide but you should because it's fun.
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[Creative Commons Attribution-NonCommercial-ShareAlike 4.0](LICENSE)
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# Always a Risk of Contamination
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Contamination occurs when other microorganisms take advantage of the gratuity you provide for your mycelium.
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There is always a risk that bacteria and mold will invade, competing with your precious mushroom makers.
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Many guides do everything they can to reduce this risk.
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My approach gives you methods that are still effective but easy and cheap to recover from contamination.
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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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- Make at least two liquid cultures for each sample we wish to test.
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- Make at least two colonization jars for each sample we wish to colonize.
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- Make one extra liquid culture and colonization jar as a control for testing your sanitation.
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Bird seed, coconut husk, and sugar water are inexpensive.
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All other equipment can be reused.
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Saving a control 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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`Spawn medium` refers to any nutrient rich growing surface, such as seed grain or simple sugar.
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# Finding Samples
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- [ ] mycelium sample
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Each section starts with a checklist of what to buy.
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The end of this document has a combined checklist.
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Newbies should start with grocery store mushrooms.
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This will allow you to walk through the methods without fear of wasting a sample.
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Finding other kinds of mycelium samples is not particularly difficult but it is frustrating to discover all the sanitary precautions were made in vain when a contaminated sample was the source.
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There remains the risk that grocery store mushrooms are contaminated.
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Online shopping can get you all sorts of mycelium 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 your harvested mushrooms.
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# Three Stages
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There are three general stages to cultivating mycelium.
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1. [Culturing](#mycelium-cultures), [wiki](https://en.wikipedia.org/wiki/Microbiological_culture)
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- Using simple sugar solution to test a sample of 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 seed grain to promote the growth of 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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The critical time to practice good sanitation is when opening any of your containers and interacting with the spawn medium inside.
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Clean yourself and work area before working with a mycelium sample or spawn medium.
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All surfaces and tools should be wiped clean then sprayed with a sanitizing solution to soak for at least 2 minute prior to working.
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Purchase a new spray bottle that is [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 during the fruiting stage.
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If you are working near your living space then a food safe sanitizer like [Star San](https://fivestarchemicals.com/star-san-sanitizer-4-oz) with avoid stinking up the place or contaminating food.
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It is very concentrate, a couple drops into a spray bottle full of water is 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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If you have a way to vent your workspace or won't bother anyone else with the fumes you can use 70% isopropyl alcohol instead of star san.
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Your face, hands, and tools are the major source of contamination.
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Tools can be sterilize in a pressure cooker or [autoclave](#autoclave).
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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, they may be more clumsy to the point of negating any benefit they might provide.
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Face masks 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 and forearms with warm water and soap to remove anything that might fall into the spawn medium.
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Avoid touching your spawn medium directly.
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Avoid touching 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 they look pretty cool when they do.
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# Mycelium Cultures
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Any samples of mycelium are potentially contaminated.
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Even using a microscope and visually checking the entire sample is tedious, error prone, and impractical.
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A small culture will allow you to grow a test batch of your sample without for a jar of [grain spawn](#grain-spawn-and-substrate) to show signs of contamination.
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The preferred culture medium is [liquid cultures](#liquid-cultures).
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[Liquid cultures](#liquid-cultures) provide a similar micro climate as agar slants with the added benefit of being a much easier for inoculating [grain spawn](#grain-spawn-and-substrate).
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The disadvantage of liquid culture is they are a little difficult to identify contaminations.
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You may use agar slants to isolate mycelium from contamination before transferring to [liquid cultures](#liquid-cultures).
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This guide does not cover their use.
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Skipping testing on agar slants works well enough as long as you give [liquid cultures](#liquid-cultures) extra time for bacteria to become visible while suspended in water.
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# Liquid Cultures
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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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- [ ] food scale, grams
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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] 50 to 100 ml syringes with hollow needles, [autoclavable](#autoclave)
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- [ ] distilled water
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- [ ] light corn syrup
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- [ ] marbles or magnetic stirring rods
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- [ ] aluminum foil
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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Imperial | Metric
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-------- | ------
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16 oz | 450 ml
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Ingredient | Mason Jar | Ratio
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---------- | --------- | -----
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water | 300 ml | 100 ml
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simple sugar | 15 g | 5 g
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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1. Fill each 16 oz mason jar with 300 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 until boiling, about 1 minute.
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3. Add 15 grams of light corn syrup to warm water.
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- Optionally add no more than 1 gram of diammonium phosphate (dap) with urea for yeast nutrients.
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4. Insert a marble or magnetic stirring rod for stirring the solution once inoculated.
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5. Cover jar opening with aluminum foil and [pressure cook](#autoclave) for 25 minutes at 70 kPa (10 psi).
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Allow to cool to room temperature.
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6. Replace aluminum foil with mason jar lid, screwing closed tightly to prevent spilling.
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7. Store in a dark cool place until inoculation.
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8. Daily shake the mason jars daily to release carbon dioxide and oxygenate the water.
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Unscrew the cap and slightly lift the lid to allow the carbon dioxide to escape.
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## Autoclave
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[Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for pressurized steam chambers 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), you may reduce the time by 1/3 for grain spawn.
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Autoclavable tools are safe to bring to 120 celsius for extended durations without damage.
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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 to the inner pot.
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2. Insert items to be autoclave on to the steam rack.
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3. Press `Steam` or `Canning` option and set the appropriate time.
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4. Turn vent valve to close.
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5. Once time has completed allow to passively depressurize.
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# Culture Inoculation
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Once you autoclave your [liquid cultures](#liquid-cultures) 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 [cloning](#cloning) grocery store 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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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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Taking mycelium samples from mushroom tissue is called cloning.
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Piercing the mushroom tissue with a hollow needled syringe is enough to collect a sample.
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [liquid culture](#liquid-cultures).
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1. Cut the stem off the mushroom with a sanitary knife.
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2. Pull the plunger of a sanitary syringe to halfway out.
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3. Plunge a hollow needle tip of the syringe into the center along the length of the stem to collect a small sample.
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4. Gently push the plunger on the syringe to use air pressure to put the sample into an open [liquid culture](#liquid-cultures).
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5. Replace the screw cap on the [liquid culture](#liquid-cultures) and store in ambient room light at room temperature.
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6. Daily release the pressure within the [liquid culture](#liquid-cultures) container by unscrewing the cap.
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## Spore Samples
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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 then be added to a syringe of sterile water for ease in shipping.
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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 [liquid culture](#liquid-cultures).
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1. Using a lighter, heat the spore syringe needle until it glows red. Allow to cool.
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2. Place a single drop of water from a spore syringe on the [liquid culture](#liquid-cultures).
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3. Replace the screw cap on the [liquid culture](#liquid-cultures) and store in ambient room light at room temperature.
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### Spore Print
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An inoculation loop maybe useful to apply the spore print samples to a [liquid culture](#liquid-cultures).
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Alternatively use a sanitary sharp knife or scalpel to apply the spore print samples.
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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- Sanitize the outside of each [liquid culture](#liquid-cultures).
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1. Lightly scrape spores off a print delivery medium with a sanitary sharp.
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2. Heat the loop element of the inoculation loop using a lighter until it glows red.
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Allow the loop to cool.
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3. Collect spores on the loop by gently rubbing against the loose spores.
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4. Insert loop into an open [liquid culture](#liquid-cultures) and gently apply spores.
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5. Replace the screw cap on the [liquid culture](#liquid-cultures) and store in ambient room light at room temperature.
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6. Daily release the pressure within the [liquid culture](#liquid-cultures) container by unscrewing the cap.
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# Reading Liquid Cultures
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[Liquid cultures](#liquid-cultures) should show growth within the first week after inoculation and should be monitored for contamination weekly.
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Mycelium are snowy white with grey, blue, or brown tints depending on the species.
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Colors like yellow, red, green, and black usually indicate contamination.
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Dispose of the contaminated solution being mindful of any stirring rods or marbles you might have inserted.
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# Colonization
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Once the mycelium is successfully isolated in [liquid cultures](#liquid-cultures), 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 sawdust.
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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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Mushroom growers have had some success with the much quicker method of inoculating store bought sterilized whole brown rice.
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These are usually sold as:
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> Instant Microwavable Whole Grain Brown Rice
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For your first time I recommend store bought sterilized whole brown rice to learn the process without worrying about contaminated grain.
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Simply cut a small hole in a corner of the sterile container and follow the instructions for [spawn inoculation](#spawn-inoculation).
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Tape the small hole closed.
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You should, however, provide mycelium a variety of [grain spawn and substrate](#grain-spawn-and-substrate) between generations of colonies so the strain does not lose it's ability to digest different sources of nutrition.
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These general instructions are the same for most [grain spawn](#grain-spawn-and-substrate) but the most accessible source of grain spawn is `wild bird seed`.
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## Grain Spawn and Substrate
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Nutrient rich grain treated and sanitized to feed mycelium.
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Vitamin and mineral rich substrate treated and sanitized to provide mycelium a growing surface.
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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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- [ ] coconut coir
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- [ ] 8 liter, approximate, bucket and lid
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- [ ] aluminum foil
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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Imperial | Metric
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16 oz | 450 ml
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1 cup | 250 ml
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Wild bird seed is 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 sanitization.
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Soaking the bird seed will clean it up a bit and encourage the endospores to reproduce, leaving them weaker to [autoclave](#autoclave).
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Earlier it was mentioned that mycelium want a variety of [grain spawn](#grain-spawn-and-substrate) and [substrates](https://en.wikipedia.org/wiki/Substrate_(biology)) between generations of colonies so the strain does not lose it's ability to digest different sources of nutrition.
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This guide uses coconut husk (coir) mixed with [grain spawn](#grain-spawn-and-substrate) for colonization as a substrate.
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You can also add coffee grounds, sawdust, straw, and rice husks for variety.
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Take caution to only change one thing at a time between generations or you risk overwhelming the mycelium.
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Be mindful of commercial gardening coir as they are treated with [endophyte](https://en.wikipedia.org/wiki/Endophyte) to protect plant roots.
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These endophytes do not protect our mushroom roots and will compete for resources like any other contamination.
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0. [Sanitize Your Workspace](#sanitize-your-workspace).
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0. Hydrate the coconut coir brick following the packaging instructions.
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0. Mix the coconut coir and wild bird seed in the bucket at a 50/50 ratio.
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0. Fill with water until moisture is visible on the surface of the mixture and soak overnight.
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0. Split evenly to each 16 oz wide mouth mason jar, about 1 cup.
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0. Cover jar opening with aluminum foil and [pressure cook](#autoclave) for 2 hours.
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0. Lightly screw lid over the foil to secure it in place without sealing the jar and allow to cool to room temperature.
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0. 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-substrate) and [liquid culture](#liquid-cultures).
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1. Gently stir the [liquid culture](#liquid-cultures) and with a sanitary syringe extract 1 ml of [liquid culture](#liquid-cultures), replace the screw top.
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2. Inject syringe into the jar of [grain spawn](#grain-spawn-and-substrate) by carefully piercing the aluminum foil straight down along the inside 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 liquid cultures](#reading-liquid-cultures), [grain spawn](#grain-spawn-and-substrate) should show growth within the first week after inoculation 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, blue, or brown tints depending on the species.
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Colors like yellow, red, green, and black usually indicate 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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Once the grain and substrate in the jar is covered with feathery white mycelium, give it one more week before [fruiting](#fruiting).
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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 without exposing the substrate to contamination.
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The mushrooms are light sensitive, to prevent them grow inside the jar we cover the glass with aluminum foil.
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## Fruiting Chambers
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- [ ] clear plastic tubs with lids, shoebox or larger
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- [ ] hygrometers
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Once the top surface of the coconut coir has been covered by feathery white mycelium it is ready to fruit.
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0. Cover the glass mason jars with a sheet of aluminum foil, remove lid exposing the colony to the air.
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0. Place jar within a clear plastic tub with the jar on its side.
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0. Place a sanitary hygrometer onto the tub where it is visible.
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0. Once tub is full of sideways open jars, replace tub lid and store in ambient room light at room temperature.
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## Daily
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Try to maintain at least 80% humidity within the fruiting chamber.
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The water soaked coconut coir should provided more than enough moisture to keep the inside of the clear plastic tubs humid.
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It is necessary to open at least daily to provide fresh air and to clear out the carbon dioxide produced by the mycelium.
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0. If needed, gently mist the mycelium cake and surrounding coconut coir a spray bottle of water until humid.
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Each mushroom is different generally harvest mushrooms when their gills are exposed but before they dump their spores and make a mess.
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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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- [ ] distilled water
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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] 50 to 100 ml syringes with hollow needles, [autoclavable](#autoclave)
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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- [ ] light corn syrup
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- [ ] marbles or magnetic stirring rods
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- [ ] flame or lighter
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- [ ] wild bird seed with millet
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- [ ] 8 liter, approximate, bucket and lid
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- [ ] aluminum foil
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- [ ] coconut coir
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- [ ] clear plastic tubs with lids, shoebox or larger
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- [ ] hygrometers
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