How to Grow Mushrooms From Tissue Culture

Prepare Agar Gel

• 1

  Slice unpeeled potatoes, place in the pot, cover with water and cook on medium high heat until the potatoes are tender.

• 2

  Strain and place liquid from the potatoes in the Pyrex bowl. Mix in 1 oz. of agar and 2 tbsp. of sugar, using the fork. Add 1 qt. of water to the mixture.

• 3

  Press a sheet of tin foil onto the Pyrex bowl to protect the contents. Add 1 cup of water to the pressure cooker. Put the bowl of agar mixture into the pressure cooker and assemble the lid. Cook on medium heat for 20 minutes at 15 psi. Start the timer when the pressure cooker reaches 15 psi. Allow the pressure cooker to cool for an hour before opening.

Fill Test Tubes

• 4

  Arrange clean tuna and soup cans in a row as they will hold the test tubes at an angle. Pour the agar mixture into the measuring cup and fill the test tubes one-third full with the agar mixture. Apply caps loosely or use foil to cover the ends of the test tubes. Place three filled test tubes in each soup can. Add 1 cup of water to the pressure cooker. Place the cans with the test tubes into the pressure cooker, then secure the lid.

• 5

  Sterilize the test tubes in the pressure cooker at 15 psi for 25 minutes. Start the timer when the pressure cooker reaches 15 psi. Allow the pressure cooker to cool on the stovetop for an hour.

• 6

  Open the pressure cooker and remove the hot test tubes carefully, using oven mitts and place three test tubes in each tuna can to hold them at the correct slant to cool. Tighten the caps after the agar has cooled and turned into a gel. Cover each can of test tubes with tin foil to keep out dust.

Prepare Mushroom Sample

• 7

  Spray the kitchen counter with bleach diluted in water and wipe with paper towels. Work in the morning to avoid a draft and airborne dust. Work fast for the remaining steps.

• 8

  Arrange the test tubes on the clean table along with the X-Acto knife, mushrooms, lighted alcohol lamp and the fourth tin can that will be used to hold the flame-sterilized X-Acto knife blade near the flame of the alcohol lamp.

• 9

  Open a test tube carefully and place it between the middle and ring finger of your left hand if you are right-handed. Hold the cap between your pinkie and ring finger.

• 10

  Break the mushroom in half and do not let the cut side touch the table. Place the X-Acto knife blade into the flame, then cut a small piece of the inside mushroom flesh and place it into the test tube. Replace the cap quickly and place the test tube with the newly created tissue culture back with the rest. Keep the X-Acto knife blade near the flame when not in use. Repeat for the eight remaining test tubes.

• 11

  Place masking tape around the bottom of the lid and record the date and variety on it, using the Sharpie marker. Keep prepared test tubes in a dark room at temperatures between 55 and 70 degrees Fahrenheit for mycelia to grow.