Protocol
Materials
- grinding tool
- scrubber
- scalpel
- hood time
- strong forceps
- vials
- ethanol in petri dish
- vial rack
Procedure
1. surface wash one log in
a. 1% sodium hypochloride – 1:10 bleach
b. water
2. scrape piece of 1 gallery in the hood, scrape into
a. 1 ml of 1X PBS AUTOCLAVED phosphate buffer saline solution in tube
3. mortar & pestle
4. plate directly, refrigeration decreases growing success
5. actual plating of Dendroctonus samples, testing three media: chitin + antibiotics, WAY + antibiotics, YMEA + streptomycin (for fungi).
a. add 0.7 ml of distilled water to the sample
b. bead beating with larger-diameter beads (710-1180 um size, about 1/3 of vial volume)
c. plate 0.3 ml of supernatant on each of the three plate types
