Modified Promega Wizard Extraction for Barcoding Macrofungi

Add 600uL of to 1.5mL eppi tubes. One tube for each specimen you are planning an extraction for.

Place tissue from your specimens into each tube using tweezers. Utilize a piece about the size of a grain of rice. The tissue can be either fresh or dried. Label the tube with the appropriate number. Wipe the tweezers off with a Kimwipe or paper towel in between each specimen. These tubes can be stored at room temperature until they are ready to be used.

Grind the tissue in each tube using a sterile pestle.

Heat the tubes at 65°C for 0h 15m 0s.

Centrifuge the tubes for 0h 3m 0s.

Transfer the supernatant (liquid on top) to a new 1.5mL eppi tube.

Add 200µL of to the tube.

Vortex the tube for 0h 0m 20s.

Centrifuge the tube for 0h 6m 0s.

Transfer the supernatant (liquid on top) to a new 1.5mL eppi tube.

Add 600µL of 100% to the tube. This precipitates the DNA.

Centrifuge the tube for 0h 1m 0s. The DNA will now be in a pellet stuck to the bottom of the tube.

Discard the supernatant. It can just be poured out of the tube into a waste container.

Add 600µL of 70% ethanol to the tube.

Centrifuge the tube for 0h 1m 0s.

Discard the supernatant. It can just be poured directly out of the tube into a waste container.

Place the tube upside down on a Kimwipe for at least 0h 15m 0s, or until all of the ethanol has evaporated from the tube. I usually leave the tube to dry overnight.

Add 30uL of molecular water to the tube.

Your DNA template is now ready for amplification.