From 3659a2636716e2a22a9088e32009aeabf4a76aea Mon Sep 17 00:00:00 2001 From: JoYo <> Date: Mon, 10 Feb 2020 19:13:45 -0500 Subject: [PATCH] wip --- README.mdown | 48 +++++++++++++++++++++++++----------------------- 1 file changed, 25 insertions(+), 23 deletions(-) diff --git a/README.mdown b/README.mdown index 952a672..0d7af46 100644 --- a/README.mdown +++ b/README.mdown @@ -171,7 +171,7 @@ MEA | 5 g - [ ] 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. 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](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.** -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. @@ -223,54 +223,55 @@ Use the following durations as they make sense to sanitize and pasteurize. 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. -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 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). -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. ## Cloning - [ ] 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. - -The collection of a sample for cloning is relatively trivial using a syringe with a hollow needle. +Taking mycelium samples from mushroom tissue is called cloning. +Piercing the mushroom tissue with a hollow needled syringe is enough to collect a sample. 0. [Sanitize Your Workspace](#sanitize-your-workspace). - 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 - [ ] inoculation loop - - [ ] 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. 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. - -In either case, an inoculation loop is used to apply the spore samples to a [culture medium](#culture-mediums). +An inoculation loop is used to apply the spore samples to a [culture medium](#culture-mediums). ### 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). -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. @@ -307,7 +308,8 @@ In either case, an inoculation loop is used to apply the spore samples to a [cul - [ ] tweezers 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. @@ -447,7 +449,7 @@ Coconut husk (coir) as casing layer and flooring in our fruiting chamber to hold ## 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.