Primers for Fungus, sequencing and identification

In other words, you're looking for a protocol to isolate fungi? I've been out of the lab for three years now, so my help is limited here, though I did provide a protocol that will show you how to isolate fungi.

THERE'S A GREAT METHOD IM USING IT FOR FUNGAL DNA EXTRACTION

To a 1.5-mL Eppendorf tube containing 500 mL of lysis buffer (400 mM Tris-HCl [pH 8.0], 60 mM ethylene diaminetetra acetic acid [EDTA] [pH 8.0], 150 mM NaCl, 1% sodium dodecyl sulfate), a small lump of mycelia from young culture is added by using a sterile toothpick, with which the lump of mycelia is disrupted. The tube is then left at room temperature for 10 min. After adding 150 mL of potassium acetate (pH 4.8; which is made of 60 mL of 5 M potassium acetate, 11.5 mL of glacial acetic acid, and 28.5 mL of distilled water), the tube is vortexed briefly and spun at 10000 × g for 1 min. The supernatant is transferred to another 1.5-mL Eppendorf tube and centrifuged again as described above. After transferring the supernatant to a new 1.5-mL Eppendorf tube, an equal volume of isopropyl alcohol is added. The tube is mixed by inversion briefly. The tube is spun at 10000 × g for 2 min, and the supernatant is discarded. The resultant DNA pellet is washed in 300 mL of 70% ethanol. After the pellet is spun at 10000 rpm for 1 min, the supernatant is discarded. The DNA pellet is air dried and dissolved in 50 mL of deionized H2O, and 1 mL of the purified DNA is used in 25 to 50 mL of PCR mixture. The extractions were done in duplicate assays for each sample.