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README.mdown
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README.mdown
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@ -171,7 +171,7 @@ MEA | 5 g
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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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Mixing liquid culture is similar to [agar slants](#agar-slants) without needing to cool the test tubes 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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@ -203,11 +203,11 @@ simple sugar | 5 g
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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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[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), reduce the time by 1/3.
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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.
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Autoclavable tools are safe to bring to 250 celsius for extended durations without damage.
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@ -223,54 +223,55 @@ Use the following durations as they make sense to sanitize and pasteurize.
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1. Add 200 ml of water.
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2. Insert items to be autoclave.
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2. Insert items to be autoclave on to the steam rack.
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3. Press Steam option and set the appropriate time.
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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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5. Once time has completed allow to passively depressurize.
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Venting the valve early may be necessary for working with agar.
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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 grocery store mushrooms.
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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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- [ ] paring knife or scalpel
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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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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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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 [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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1. With a sanitary knife, cut the stem off the mushroom.
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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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2. Pull the plunger of a sanitary syringe to halfway out.
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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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3. Using the hollow needle tip of the syringe, collecting a sample of the core by plunging into the center along the length of the stem.
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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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4. Gently use air pressure from the syringe will push the sample into an open [culture medium](#culture-mediums).
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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 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 then be added to a syringe of sterile water for ease in shipping.
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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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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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@ -278,11 +279,11 @@ In either case, an inoculation loop is used to apply the spore samples to a [cul
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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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1. Using a lighter to heat the loop element of the 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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2. Scrape spores off a print delivery medium with a sterile sharp.
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3. Collect spores on the loop.
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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 [culture medium](#culture-mediums) and gently apply spores.
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@ -307,7 +308,8 @@ In either case, an inoculation loop is used to apply the spore samples to a [cul
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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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Usually these can be used to inoculate [grain spawn](#grain-spawn) directly.
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Cultures are used 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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@ -447,7 +449,7 @@ Coconut husk (coir) as casing layer and flooring in our fruiting chamber to hold
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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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Try to maintain 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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