diff --git a/README.mdown b/README.mdown index 8e38b2e..148a294 100644 --- a/README.mdown +++ b/README.mdown @@ -108,33 +108,68 @@ There are three general stages to cultivating mycelium. # Culturing Any samples of mycelium are potentially contaminated. -Even using a microscope and visually checking the entire sample is tedious and error prone. -A small culture will allow you to grow a test patch of your sample without risking a whole jar of [grain spawn](#grain-spawn). +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 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 givings mycelium and contamination a surface to grow in large enough colonies to view with the naked eye. +Agar gives 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). 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. +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). -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 - [ ] food scale, grams -- [ ] 16 oz wide mouth mason jars with lids - [ ] distilled water -- [ ] 50 to 100 ml syringe with hollow needle, [autoclavable](#autoclave) -- [ ] 30 to 50 ml test tube with screw cap, [autoclavable](#autoclave) +- [ ] 16 oz wide mouth mason jars with lids - [ ] 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 - [ ] 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. 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: -| | | --- | -- -water | 100 ml -MEA | 5 g +ingredient | mason jar | ratio +---------- | --------- | ----- +water | 300 ml | 100 ml +MEA | 15 g | 5 g 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). 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. @@ -171,40 +206,6 @@ MEA | 5 g 8. Store in a dark cool place until inoculation. 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](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. -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.