Mar 27, 2024

Public workspaceDrosophila melanogaster inoculation with Bd V.1

DOI
https://dx.doi.org/10.17504/protocols.io.yxmvme7n6g3p/v1
  • Tatum Katz1,
  • Alyssa Byer2,
  • Renwei Chen2
  • 1US Meat Animal Research Center, Agricultural Research Service, US Department of Agriculture, Clay Center, NE, USA;
  • 2University of California, Santa Barbara
Tatum S Katz
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DOI: https://dx.doi.org/10.17504/protocols.io.yxmvme7n6g3p/v1
Protocol Citation: Tatum Katz, Alyssa Byer, Renwei Chen 2024. Drosophila melanogaster inoculation with Bd. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvme7n6g3p/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 27, 2024
Last Modified: March 27, 2024
Protocol Integer ID: 97460
Keywords: drosophila melanogaster inoculation with bd method, drosophila melanogaster inoculation, batrachochytrium dendrobatidi, batrachochytrium dendrobatidis under review, manuscript drosophila melanogaster, bd method
Funders Acknowledgements:
NIH Ecology and Evolution of Infectious Diseases
Grant ID: R01GM135935
University of California Santa Barbara California Alliance for Minority Participation
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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.
Troubleshooting
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.