downplay agar slants, a likely unessisary step.
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README.mdown
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README.mdown
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@ -108,33 +108,68 @@ There are three general stages to cultivating mycelium.
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# Culturing
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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 and error prone.
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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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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) to show signs of contamination.
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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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Agar givings mycelium and contamination a surface to grow in large enough colonies to view with the naked eye.
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Agar gives mycelium and contamination a surface to grow in large enough colonies to view with the naked eye.
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A week of growth is usually long enough to produce colonies which can be identified and transferred to [liquid cultures](#liquid-cultures).
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While petri dishes are part of the usual trappings of cultivating microorganisms, they are fairly high maintenance and better suited for a laboratory environment.
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[Agar slants](#agar-slants) provide the same functionality without the need to seal and re-seal with parafilm.
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The preferred [culture medium](#culture-mediums) is [liquid cultures](#liquid-cultures).
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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 they are difficult to identify contaminations.
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The disadvantage of liquid culture is they are a little difficult to identify contaminations.
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You should use [agar slants](#agar-slants) to isolate mycelium from contamination into [liquid cultures](#liquid-cultures).
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You may use [agar slants](#agar-slants) to isolate mycelium from contamination before transferring to [liquid cultures](#liquid-cultures).
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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.
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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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- [ ] 16 oz wide mouth mason jars with lids
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- [ ] electric pressure cooker, stove top, or [autoclave](#autoclave)
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- [ ] aluminum foil
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imperial | metric
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-------- | ------
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16 oz | 450 ml
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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 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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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.
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3. Add 15 grams of light corn syrup to warm water and stir for 1 minute with a sanitary utensil.
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4. Cover un-lid jar opening with aluminum foil and [autoclave](#autoclave) for 1 hour.
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5. Allow to cool and replace aluminum foil with mason jar lid.
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6. Store in a dark cool place until inoculation.
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7. Once inoculated, shake the mason jars daily to oxygenate the water and break up the mycelium.
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## Agar Slants
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- [ ] malt extract agar
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- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave)
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- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave)
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Agar agar is a plant gelatin that solidifies at room temperature, providing a physical surface that mycelium to 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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@ -143,20 +178,20 @@ Mixing 2:3 grams ratio simple sugar to Agar powder is good for saving money in b
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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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-- | --
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water | 100 ml
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MEA | 5 g
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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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MEA | 15 g | 5 g
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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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1. Fill a 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.
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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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3. Add 15 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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@ -171,40 +206,6 @@ MEA | 5 g
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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 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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-- | --
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water | 100 ml
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simple sugar | 5 g
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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 until 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 pressurized steam chambers for sanitization and sterilization.
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@ -225,7 +226,7 @@ Use the following durations as they make sense to sanitize and pasteurize.
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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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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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