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@ -171,7 +171,7 @@ MEA | 5 g
- [ ] light corn syrup - [ ] light corn syrup
Mixing liquid culture is similar to [agar slants](#agar-slants) without needing to cool in any particular position. Mixing liquid culture is similar to [agar slants](#agar-slants) without needing to cool the test tubes in any particular position.
A simple sugar like light corn syrup is dissolved in distilled water to provide mycelium calories for reproduction. A simple sugar like light corn syrup is dissolved in distilled water to provide mycelium calories for reproduction.
Scale the following recipe as needed to fill 2/3 of each test tube or 20 ml: Scale the following recipe as needed to fill 2/3 of each test tube or 20 ml:
@ -203,11 +203,11 @@ simple sugar | 5 g
## Autoclave ## Autoclave
[Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for a pressurized steam chamber for sanitization and sterilization. [Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for pressurized steam chambers for sanitization and sterilization.
**All autoclave duration measurements use electric pressure cookers for convenience.** **All autoclave duration measurements use electric pressure cookers for convenience.**
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. 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.
Autoclavable tools are safe to bring to 250 celsius for extended durations without damage. Autoclavable tools are safe to bring to 250 celsius for extended durations without damage.
@ -223,54 +223,55 @@ Use the following durations as they make sense to sanitize and pasteurize.
1. Add 200 ml of water. 1. Add 200 ml of water.
2. Insert items to be autoclave. 2. Insert items to be autoclave on to the steam rack.
3. Press Steam option and set the appropriate time. 3. Press `Steam` option and set the appropriate time.
4. Turn vent valve to close. 4. Turn vent valve to close.
5. Quick release once time has completed if needed, working with agar warm is easier. 5. Once time has completed allow to passively depressurize.
Venting the valve early may be necessary for working with agar.
# Culture Inoculation # Culture Inoculation
Once you autoclave your [culture mediums](#culture-mediums) and they have cooled to room temperature it is ready for mycelium. Once you autoclave your [culture mediums](#culture-mediums) and they have cooled to room temperature it is ready for mycelium.
Depending on the source of the sample there are two ways to inoculate, [cloning](#cloning) from a mushroom or using [spore samples](spore-samples). Depending on the source of the sample there are two ways to inoculate, [cloning](#cloning) from a mushroom or using [spore samples](spore-samples).
Newbies should start with grocery store mushrooms. Newbies should start with [cloning](#cloning) grocery store mushrooms.
This will allow you to walk through the methods without fear of wasting a spore sample. This will allow you to walk through the methods without fear of wasting a spore sample.
## Cloning ## Cloning
- [ ] paring knife or scalpel - [ ] paring knife or scalpel
- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
Taking a mycelium samples from the stem of a mushroom is called cloning. Taking mycelium samples from mushroom tissue is called cloning.
Piercing the mushroom tissue with a hollow needled syringe is enough to collect a sample.
The collection of a sample for cloning is relatively trivial using a syringe with a hollow needle.
0. [Sanitize Your Workspace](#sanitize-your-workspace). 0. [Sanitize Your Workspace](#sanitize-your-workspace).
- Sanitize the outside of each [culture medium](#culture-mediums). - Sanitize the outside of each [culture medium](#culture-mediums).
1. With a sanitary knife, cut the stem off the mushroom then half down the length. 1. With a sanitary knife, cut the stem off the mushroom.
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. 2. Pull the plunger of a sanitary syringe to halfway out.
3. Gently use air pressure from the syringe will push the sample into an open [culture medium](#culture-mediums). 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.
4. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature. 4. Gently use air pressure from the syringe will push the sample into an open [culture medium](#culture-mediums).
5. Replace the screw cap on the [culture medium](#culture-mediums) and store in ambient room light at room temperature.
## Spore Samples ## Spore Samples
- [ ] inoculation loop - [ ] inoculation loop
- [ ] flame or lighter - [ ] flame or lighter
[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. [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.
These are called spore prints. These are called spore prints.
Spores can then be added to a syringe of sterile water for ease in shipping.
Spores can further be added to a syringe of sterile water for ease in shipping. An inoculation loop is used to apply the spore samples to a [culture medium](#culture-mediums).
In either case, an inoculation loop is used to apply the spore samples to a [culture medium](#culture-mediums).
### Spore Print ### Spore Print
@ -278,11 +279,11 @@ In either case, an inoculation loop is used to apply the spore samples to a [cul
- Sanitize the outside of each [culture medium](#culture-mediums). - Sanitize the outside of each [culture medium](#culture-mediums).
1. Using a lighter, heat the loop element of your inoculation loop until it glows red. Allow the loop to cool. 1. Using a lighter to heat the loop element of the inoculation loop until it glows red. Allow the loop to cool.
2. Scraped spores off a print delivery medium with a sterile sharp. 2. Scrape spores off a print delivery medium with a sterile sharp.
3. Collect spores on the loop. 3. Collect spores on the loop by gently rubbing against the loose spores.
4. Insert loop into an open [culture medium](#culture-mediums) and gently apply spores. 4. Insert loop into an open [culture medium](#culture-mediums) and gently apply spores.
@ -307,7 +308,8 @@ In either case, an inoculation loop is used to apply the spore samples to a [cul
- [ ] tweezers - [ ] tweezers
Mycelium samples are often sold as colonized grain or wood plugs. Mycelium samples are often sold as colonized grain or wood plugs.
Usually these can be used to inoculate [grain spawn](#grain-spawn) directly but we can also make cultures to store these samples long-term. Usually these can be used to inoculate [grain spawn](#grain-spawn) directly.
Cultures are used to store these samples long-term.
With sanitary tweezers place a sample in an [agar slant](#agar-slants), [liquid culture](#liquid-cultures), or grain spawn. With sanitary tweezers place a sample in an [agar slant](#agar-slants), [liquid culture](#liquid-cultures), or grain spawn.
@ -447,7 +449,7 @@ Coconut husk (coir) as casing layer and flooring in our fruiting chamber to hold
## Daily ## Daily
The goal is to maintain a 90% humidity within the fruiting chamber. Try to maintain 90% humidity within the fruiting chamber.
0. With a water spray bottle gently mist the coconut coir and mycelium cake until the coir is moist but not soaked. 0. With a water spray bottle gently mist the coconut coir and mycelium cake until the coir is moist but not soaked.