Materials and Reagents

1. Standard materials and reagents
   1. Sterile microcentrifuge tubes, 1.5 or 2.0 ml
   2. Micropipette tips: 10, 200, 1,000 µl
   3. Miracloth (Calbiochem, e.g., VWR, catalog number: 475855-1 )
   4. Plant or fungal samples
   5. Purified (RO, DI, MilliQ, Nanopure) and sterilized water
   6. 100 bp DNA ladder (Jena Bioscience, catalog number: M-214S )
   7. 1 kb DNA ladder (New England Biolabs, catalog number: N0552 )
   8. NaCl
   9. KCl
   10. EDTA
   11. SDS
   12. Tris (pH 7.5)
   13. Polyvinypyrrolidone (PVP, CAS 900-39-8, FW 40,000, e.g., Caisson Labs, catalog number: P071-100GM )
   14. Isopropanol
   15. Ethanol (EtOH)
   16. Extraction buffer (see Recipes)
   17. Wash buffer (see Recipes)
       
       
2. Special materials and reagents for different tissue homogenization optionsB.Special materials and reagents for different tissue homogenization options
   1. Option 1 for soft leaf tissue and mycelium: No special materials or reagents
   2. Option 2 for various plant tissue and mycelium, larger than 100 mg: Liquid nitrogen
   3. Option 3 for various plant tissue and mycelium, smaller than 100 mg: Bead beater tubes (e.g., Lysing Matrix I, MP Biomedicals, catalog number: 116918050-CF )
      Note: To reduce plastic waste and save on costs, the bead beater tubes can be washed and re-used (see Notes).
      

Equipment

1. Standard equipment
   1. Scalpels
   2. Tweezers
   3. Spatulas
   4. Scale (e.g., Metler-Toledo, model: ML54T )
   5. Micropipettes: 1, 10, 100, 1,000 µl
   6. Heat block (e.g., VWR, catalog number: 12621-096 )
   7. Vortex (e.g., VWR, catalog number: 10153-838 )
   8. Microcentrifuge (e.g., Eppendorf, model: 5424 )
   9. One of the following to heat up water: Microwave, waterbath (e.g., VWR, model: WB05 ), or hot plate (e.g., VWR, catalog number: NO97042-642 )
   10. Freezer, -20 °C or -80 °C
       
       
2. Special equipment for different tissue homogenization options
   1. Option 1 for soft leaf tissue and mycelium: No special equipment
   2. Option 2 for various plant tissue and mycelium, more than 100 mg:
      Dewar for liquid nitrogen
      Ceramic mortars and pestles
   3. Option 3 for various plant tissue and mycelium, less than 100 mg:
      Bead beater (e.g., Biospec, model: Mini-Beadbeater 16 , catalog number: 607/607EUR )
   4. For sampling xylem tissue from mature trees: chisel (e.g., Grainger, catalog number: 2AJA6 ) and mallet (e.g., Grainger, catalog number: 4YR61 ), cutting board
      

Procedure