Drosophila melanogaster inoculation with Bd
Tatum S Katz, Alyssa Byer, Renwei Chen
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Abstract
Methods to inoculate D. melanogaster with Batrachochytrium dendrobatidis .
For manuscript Drosophila melanogaster is a possible vector of Batrachochytrium dendrobatidis under review at PLOS One.
Steps
Protocol
Plate 0.5ml of treatment (10^6 zsps per fly or DI water) onto each agar plate
Allow plates to dry so that no water is puddled on the surface
Place 10 flies onto each plate
Seal plates with parafilm and incubate at room temperature for 24 hours
Transfer flies into fly vials (keeping all flies from the same plate together in the same vial)
Euthanize flies in the freezer at desired time point (within 5 days). Or, use the living inoculated flies for another protocol
If you want to test the difference between washed and unwashed flies, you may wash a group of up to 5 flies at a time in PBS buffer using 500ul per fly prior to DNA extraction.
Inoculated flies are now ready for DNA extraction or other methods.
