Corex centrifuge tube

Fungal mycelia was collected by vacuum filtration using a Büchner funnel and Whatman® qualitative filter paper. The mycelium was disrupted by grinding with mortar and pestle with the addition of 2 mL of isolation buffer [10 mM Tris-HCl (pH 8.0), 440 mM sucrose, 5 mM ethylene-diamine-tetra-acetic acid (EDTA)] and 1.5 g of acid-washed sand for each 1 g of mycelia. The fungal material was ground for about 5–10 minutes until the mycelia/sand/buffer mixtures forms a slurry. This slurry was transferred to a 25 mL Corex® centrifuge tube (ThermoFisher) and centrifuged for 15 min at 3000 g using a Sorvall® SS-34 fixed angle rotor in a Sorvall® RC-5B Plus centrifuge at 4 °C to pellet nuclei, cell debris and sand. The supernatant was transferred to a 25 mL Corex® centrifuge tube (ThermoFisher) and centrifuged at 20 000 g using a Sorvall® SS-34 fixed angle rotor in a Sorvall® RC-5B Plus centrifuge at 4 °C for 30 min to pellet the mitochondria.