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downplay agar slants, a likely unessisary step.

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JoYo 2020-02-15 16:01:20 -05:00
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@ -108,33 +108,68 @@ There are three general stages to cultivating mycelium.
# Culturing # Culturing
Any samples of mycelium are potentially contaminated. Any samples of mycelium are potentially contaminated.
Even using a microscope and visually checking the entire sample is tedious and error prone. Even using a microscope and visually checking the entire sample is tedious, error prone, and impractical.
A small culture will allow you to grow a test patch of your sample without risking a whole jar of [grain spawn](#grain-spawn). A small culture will allow you to grow a test batch of your sample without for a jar of [grain spawn](#grain-spawn) to show signs of contamination.
The two [culture mediums](#culture-mediums) preferred are [agar slants](#agar-slants) and [liquid cultures](#liquid-cultures) together. Agar gives mycelium and contamination a surface to grow in large enough colonies to view with the naked eye.
Agar givings mycelium and contamination a surface to grow in large enough colonies to view with the naked eye.
A week of growth is usually long enough to produce colonies which can be identified and transferred to [liquid cultures](#liquid-cultures). A week of growth is usually long enough to produce colonies which can be identified and transferred to [liquid cultures](#liquid-cultures).
While petri dishes are part of the usual trappings of cultivating microorganisms, they are fairly high maintenance and better suited for a laboratory environment. While petri dishes are part of the usual trappings of cultivating microorganisms, they are fairly high maintenance and better suited for a laboratory environment.
[Agar slants](#agar-slants) provide the same functionality without the need to seal and re-seal with parafilm. [Agar slants](#agar-slants) provide the same functionality without the need to seal and re-seal with parafilm.
The preferred [culture medium](#culture-mediums) is [liquid cultures](#liquid-cultures).
[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). [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).
The disadvantage of liquid culture is they are difficult to identify contaminations. The disadvantage of liquid culture is they are a little difficult to identify contaminations.
You should use [agar slants](#agar-slants) to isolate mycelium from contamination into [liquid cultures](#liquid-cultures). You may use [agar slants](#agar-slants) to isolate mycelium from contamination before transferring to [liquid cultures](#liquid-cultures).
Skipping testing on [agar slants](#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.
# Culture Mediums # Culture Mediums
- [ ] food scale, grams - [ ] food scale, grams
- [ ] 16 oz wide mouth mason jars with lids
- [ ] distilled water - [ ] distilled water
- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave) - [ ] 16 oz wide mouth mason jars with lids
- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave)
- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave) - [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
- [ ] aluminum foil
imperial | metric
-------- | ------
16 oz | 450 ml
## Liquid Cultures
- [ ] light corn syrup
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.
ingredient | mason jar | ratio
---------- | --------- | -----
water | 300 ml | 100 ml
simple sugar | 15 g | 5 g
0. [Sanitize Your Workspace](#sanitize-your-workspace).
1. Fill each 16 oz mason jar with 300 ml water on a food scale.
- Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure).
2. Microwave with a safe cover until boiling.
3. Add 15 grams of light corn syrup to warm water and stir for 1 minute with a sanitary utensil.
4. Cover un-lid jar opening with aluminum foil and [autoclave](#autoclave) for 1 hour.
5. Allow to cool and replace aluminum foil with mason jar lid.
6. Store in a dark cool place until inoculation.
7. Once inoculated, shake the mason jars daily to oxygenate the water and break up the mycelium.
## Agar Slants ## Agar Slants
- [ ] malt extract agar - [ ] malt extract agar
- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave)
- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
Agar agar is a plant gelatin that solidifies at room temperature, providing a physical surface that mycelium to colonize. Agar agar is a plant gelatin that solidifies at room temperature, providing a physical surface that mycelium to colonize.
This is mixed with a simple sugar like malt extract to provide mycelium calories for reproduction. This is mixed with a simple sugar like malt extract to provide mycelium calories for reproduction.
@ -143,20 +178,20 @@ Mixing 2:3 grams ratio simple sugar to Agar powder is good for saving money in b
Scale the following recipe as needed to fill 1/3 of each test tube or 10 ml: Scale the following recipe as needed to fill 1/3 of each test tube or 10 ml:
| | | ingredient | mason jar | ratio
-- | -- ---------- | --------- | -----
water | 100 ml water | 300 ml | 100 ml
MEA | 5 g MEA | 15 g | 5 g
0. [Sanitize Your Workspace](#sanitize-your-workspace). 0. [Sanitize Your Workspace](#sanitize-your-workspace).
1. Fill a 16 oz mason jar with 100 ml water on a food scale. 1. Fill a 16 oz mason jar with 300 ml water on a food scale.
- Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure). - Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure).
2. Microwave with a safe cover until boiling. 2. Microwave with a safe cover until boiling.
3. Add 5 grams of MEA to warm water and stir for 1 minute with a sanitary utensil. 3. Add 15 grams of MEA to warm water and stir for 1 minute with a sanitary utensil.
4. While the MEA solution is still warm, fill a sanitary syringe with the solution. 4. While the MEA solution is still warm, fill a sanitary syringe with the solution.
@ -171,40 +206,6 @@ MEA | 5 g
8. Store in a dark cool place until inoculation. 8. Store in a dark cool place until inoculation.
Do not freeze. Do not freeze.
## Liquid Cultures
- [ ] light corn syrup
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.
Scale the following recipe as needed to fill 2/3 of each test tube or 20 ml:
| | |
-- | --
water | 100 ml
simple sugar | 5 g
0. [Sanitize Your Workspace](#sanitize-your-workspace).
1. Fill a 16 oz mason jar with 100 ml water on a food scale.
- Grams is the same as milliliters of water at [STP](https://en.wikipedia.org/wiki/Standard_conditions_for_temperature_and_pressure).
2. Microwave with a safe cover until boiling.
3. Add 5 grams of light corn syrup to warm water and stir for 1 minute with a sanitary utensil.
4. Fill a sanitary syringe with the solution.
5. Fill a sanitary test tube one third (2/3) capacity and replace the screw cap.
6. Place test tubes in an empty mason jar and [autoclave](#autoclave) for 1 hour.
7. Store in a dark cool place until inoculation.
8. Once inoculated, shake the test tubes daily to oxygenate the water and break up the mycelium.
## Autoclave ## Autoclave
[Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for pressurized steam chambers for sanitization and sterilization. [Autoclave](https://en.wikipedia.org/wiki/Autoclave) is a generic term for pressurized steam chambers for sanitization and sterilization.
@ -225,7 +226,7 @@ Use the following durations as they make sense to sanitize and pasteurize.
0. Add the steam rack to the inner pot of the electric pressure cooker. 0. Add the steam rack to the inner pot of the electric pressure cooker.
1. Add 200 ml of water. 1. Add 200 ml of water to the inner pot.
2. Insert items to be autoclave on to the steam rack. 2. Insert items to be autoclave on to the steam rack.