User Tools

Site Tools


sop_rye_grain_spawn

Differences

This shows you the differences between two versions of the page.

Link to this comparison view

Next revision
Previous revision
sop_rye_grain_spawn [2020/10/23 01:05]
mwikiadmin created
sop_rye_grain_spawn [2020/10/23 14:14] (current)
mwikiadmin
Line 2: Line 2:
 This SOP is adapted from Penn State Spawn Lab Procedures ((https://​plantpath.psu.edu/​about/​facilities/​mushroom/​cultures-spawn/​spawn-preparation)) for the home user who may not have access to a laminar flow hoods, Erlenmeyer flasks, Autoclaves, etc. This SOP is adapted from Penn State Spawn Lab Procedures ((https://​plantpath.psu.edu/​about/​facilities/​mushroom/​cultures-spawn/​spawn-preparation)) for the home user who may not have access to a laminar flow hoods, Erlenmeyer flasks, Autoclaves, etc.
  
-**RYE GRAIN SPAWN**+**__RYE GRAIN SPAWN__**
  
  
 **Step 1** Prepare a container for the pressure cooker, either: **Step 1** Prepare a container for the pressure cooker, either:
  
-(1) 8oz Wide Mouth Canning Jar with lid modified with a self-healing inoculation port and fresh-air-exchange port; or (2)Use a small mushroom autoclave-safe grow bag with an innoculation port and fresh-air-exchange port.+(1) 8oz Wide Mouth Canning Jar with lid modified with a self-healing inoculation port and fresh-air-exchange port; or 
  
-**Step 2** Measure out the ingredients +(2)Use a small mushroom autoclave-safe grow bag with an innoculation port and fresh-air-exchange port. 
-ORGANIC RYE WHOLE GRAIN - Using a measuring cup, about 50 ml (or if using a kitchen scale, about 6 ounces or 170grams ​+ 
-LIME POWDER (Calcium sulfate) - 1/2 teaspoon Calcium Carbonate +**Step 2** Measure out the three ingredients 
-GYPSUM POWDER ​ (Calcium sulfate)- 1/4 teaspoon +(1) ORGANIC RYE WHOLE GRAIN - Using a measuring cup, about 50 ml (or if using a kitchen scale, about 30 grams
-WARM WATER - 60mL (distilled water preferred but not required)+(2)LIME POWDER (Calcium sulfate) - 1/2 teaspoon Calcium Carbonate 
 +(3) GYPSUM POWDER ​ (Calcium sulfate)- 1/4 teaspoon 
 +WARM WATER - 60mL (distilled water preferred but not required. Any water about ph7 will be fine.)
  
 **Step 3** Prepare the bag or jar for sterilization. **Step 3** Prepare the bag or jar for sterilization.
 Add all ingredients and seal your canning jar or grow bag, and mix the ingredients gently. Add all ingredients and seal your canning jar or grow bag, and mix the ingredients gently.
  
-If using a canning jar, make sure to twist the ring hand tight. It should be airtight and there is no concern of over pressurizing ​the jar or having it hold vacuum because of the fresh-air-exchange port that allows the jar to breath. Put a piece of foil over the top of the jar so that you can minimize the risk of contaminating the top of the lid from when you transfer it out of the pressure cooker into your ((Still Air Box)) or ((Laminar flow hood)) for inoculation. ​+If using a canning jar, make sure to twist the ring hand tight. It should be airtight and there is no concern of overpressurizing ​the jar or having it hold vacuum because of the fresh-air-exchange port that allows the jar to breath. Put a piece of foil over the top of the jar so that you can minimize the risk of contaminating the top of the lid from when you transfer it out of the pressure cooker into your [[still_air_box|still air box]] or [[laminar_flow_hood|Laminar flow hood]] for inoculation. ​
  
 If using a mushroom bag, try to push out as much air as possible, and seal the top with binder clips, a hair iron, rubber band, or even tie the top off in a nice tight knot.  If using a mushroom bag, try to push out as much air as possible, and seal the top with binder clips, a hair iron, rubber band, or even tie the top off in a nice tight knot. 
  
-*Step 4* Sterilize the grain+**Step 4** Sterilize the grain
 Place the jar(s) or bag(s) into pressure cooker, and maintain 15 psi for at least 90 minutes for smaller batches. Bigger batches will take longer. It is theoretically possible to sterilize for too long, you will know it is too long if the seeds have exploded out of their hulls. ​ Place the jar(s) or bag(s) into pressure cooker, and maintain 15 psi for at least 90 minutes for smaller batches. Bigger batches will take longer. It is theoretically possible to sterilize for too long, you will know it is too long if the seeds have exploded out of their hulls. ​
  
-*Step 5* Inoculate the sterilized grain + 
-Allow the sterilized grain to cool to room temperature. Transfer the sterilized jar or bag to your ((Still Air Box)) or ((Laminar ​Flow Hood)) ​and using a sterile syringe tip, inject your liquid culture through the self-healing inoculation port.  +**Step 5** Inoculate the sterilized grain 
-Usually, I use an ((alcohol burner)) to get the steel syringe tip wet hot, cool it slightly with a single-use alcohol wipe, squirt out a few drops to clear the barrel of any microbials that might have survived the flame and alcohol, before injecting into the sterilized grain. ​+Allow the sterilized grain to cool to room temperature. Transfer the sterilized jar or bag to your [[still_air_box|still air box]] or [[laminar_flow_hood|Laminar ​flow hood]]and using a sterile syringe tip, inject your liquid culture through the self-healing inoculation port.  
 +Usually, I use an alcohol burner((alcohol_burner)) to get the steel syringe tip wet hot, cool it slightly with a single-use alcohol wipe, squirt out a few drops to clear the barrel of any microbials that might have survived the flame and alcohol, before injecting into the sterilized grain. ​
  
 If your syringe tip is long enough, try to distribute the liquid culture in different spots of the grain so the entire mass will colonize more quickly. If your syringe tip is long enough, try to distribute the liquid culture in different spots of the grain so the entire mass will colonize more quickly.
  
-*Step 6* Wait for colonization +**Step 6** Wait for colonization 
-Depending on your ambient conditions and your particular cultivar , you should begin to see fuzzy white mycelium growth within 2-4 weeks. The more fully colonized the grain is, the more success you will have when you ((transplant)) the colonized grain to your grow substrate. ​+Depending on your ambient conditions and your particular cultivar , you should begin to see fuzzy white mycelium growth within 2-4 weeks. The more fully colonized the grain is, the more success you will have when you transplant((transplant)) the colonized grain to your grow substrate. ​
  
sop_rye_grain_spawn.1603429535.txt.gz · Last modified: 2020/10/23 01:05 by mwikiadmin