Hi all,
For the transformation of Botrytis, I am starting with a spore suspension from a week-10 day old petri dish. Initially I usually get a count of around 5,00,000 spores/ml. I encounter problem when I have to inoculate this spore suspension in two 50 ml PDB flasks.
The final spore concentration in 100 ml PDB has to be 10^6/ml. What method should I adopt? Should I add the entire spore suspension (of around 5 ml), into the flask or should I centrifuge the suspension and get a pellet to concentrate the spore suspension?
Inoculum suspension inoculum hasn't the titer to establish 10E6/ml in 50 ml .
Try centrifugation and resuspension in a small volume. You prob should do that anyway to have a clean prep. In that you'll need to ensure effective resuspension - maybe add a little Tween. However, even for this, the initial volume may be problematic as you'll need 10 fold+ in crease in titer.
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