The purpose of this experiment is to determine if P. larvae is able to germinate in the Bee Artificial Diet (BAD).
Used the ten fold serially diluted Spore Stock A that was created on 6-27-14 that had since been stored at 4C. Added 100 uL of the 10^-1 to 400 uL of BAD in eppendorf tubes. Repeated for dilutions 10^-2 to 10^-4.
For controls, added 100 uL of each dilution to 400 uL of BHI+thiamine and 100 uL of each dilution to 400 uL of H2O.
Incubated all samples at 35C shaking at 150 rpm for 2.5 hours.
Performed drop plate procedure per Link for each dilution under each treatment onto MYPGP agar. (note: 10 uL per drop, repeated five times). Incubated plates not inverted at 35C 5% CO2. Remaining samples were stored at 4C.
I am not sure how many colonies I will actually be able to identify in the BAD treatment group, as the media is fairly cream colored and slightly viscous.
Perhaps I should have made dilutions from each sample, but since they were already diluted before being diluted even further with different treatments and only incubated for 2.5 hours, it may not be necessary...
//EWW
No comments:
Post a Comment