From 5-4-15.
Two isolation streaks of P. larvae 9545 (from the -80C stock) were struck out onto MYPGP agar plates and incubated at 37C for three days until nice isolated colonies formed. A single isolated colony was inoculated into 10 mL of BHI broth and incubated at 37C 225 rpm for 24 hours.
I made more Columbian blood agar plates (Columbian agar base + 5% defibrinated sheep's blood). I am currently out of fresh defibrinated sheep's blood. I made plates and not slants this time since it appears that I can recover a similar concentration of spores, if not a higher titer, from the plates. Plus, I was having issues with using the slants with accidentally removing agar from them and transferring them to the Eppendorf tubes during the spore extraction procedure.
A 100 uL volume of the overnight culture was inoculated and spread onto the blood agar plates. The plates were incubated at 37C 5%CO2 in the media kitchen (Van Es Room 115) incubator was used. The plates will incubate for seven days until the spores will be extracted and purified.
//EWW
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