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Home» Bacteria » Extraction from Galleries

Extraction from Galleries

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

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