STEP BY STEP GUIDE ON HOW TO PERFORM TISSUE CULTURE

 

Prepare materials

  1. Scalpel or knife with insulated handle
  2. Forcep, curved or pointed
  3. Isopropyl alcohol 70%
  4. Hand towel
  5. Wet wipe
  6. Lab burner
  7. Lighter
  8. Fresh oyster mushroom
  9. Containers filled with PDA
  10. Face mask
  11. Surgical hand glove
  12.  Label materials or Marker (optional)

 

Select a strong mushroom for culture

  1. Healthy
  2. Not too mature, not too young
  3. Not too humid (preferably after 2-3 hours of watering)
  4. With a broad stipe/stalk
  5. Make sure it is clean and far from any contaminated mushroom

 

How to

  1. Clean the work area with a wet wipe moist with alcohol
  2. Clean all necessary tools with a wet wipe moist with alcohol
  3. Then heat it over the fire until RED hot
  4. Place all cleaned materials aside and allow it cool down for 5 – 10 minutes
  5. Clean both hands with alcohol and put on the glove
  6. Clean the PDA containers with a wet wipe moist with alcohol
  7. Heat scalpel or knife until turning RED hot. Make sure it does not touch any surface after heating
  8. Alternatively, dip the tools in a cup of alcohol to cool it down
  9. Use fingers to tear stalk lengthwise or use the scalpel
  10. With the forceps, gently tear a small piece of fleshy tissue from inside the stalk
  11. Make sure it is clean and did not touch the outside of the mushroom
  12. Uncover the PDA container
  13. Hover the forceps above the container and inoculate by placing a small piece of mushroom tissue in the middle of the PDA surface
  14. Make sure the piece of  mushroom tissue does not touch anything before the entrance
  15. Cover the containers immediately and wipe them down with alcohol
  16. Label them with the date and species type
  17. The time of incubation to full growth mycelium will take about 10 – 15 days (depending on species)

 

Caring Process

  1. Keep the PDA tissue culture on a clean shelf
  2. Check the contamination and growth rate daily
  3. After mycelium covers the whole PDA medium, keep mycelium in a cool place or refrigerator
  4. Separate contaminated PDA container away from good ones
  5. Keep a detailed record of observation

 

 

 

 

Recommend Online Courses

STEP BY STEP GUIDE ON HOW TO PREPARE POTATO DEXTROSE AGAR

STEP BY STEP GUIDE ON HOW TO PREPARE GRAINS

STEP BY STEP GUIDE ON HOW TO PREPARE GRAINS FOR INOCULATION

STEP BY STEP GUIDE ON HOW TO PREPARE SUBSTRATE

STEP BY STEP GUIDE ON HOW TO INOCULATE THE SUBSTRATE

STEP BY STEP GUIDE ON HOW TO MANAGE THE DEVELOPMENT OF MUSHROOM BAG

 

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