I have been screening medicinal plant extracts against C. neoformans on 96 well plates (MIC assays) for 6 months. Lately I have been having trouble with slow growing yeast. I add INT dye to detect for growth after 3 days (72 hours) then I would take the MIC value 19 hours later when there is enough growth in my growth control.
Lately however - the yeast has not been growing well on a proportion of my assay plates by the time I take my MIC values - and they were definitely prepared from the same batch and the experiments were completed in exactly the same way - I have no idea what is happening - any ideas from anyone? I do not work from the Stock culture every time - but from the previous week's culture plates - could it be that my yeast has changed?
Hi Chris,
I have been doing similar experiments with Cryptococcus. I have faced problems with a general reduction in growth once and it got resolved when I went back to my main stock in deep freezer. I would suggest you to go back to your main stock (glycerol stock) and do the experiment if it a general issue. It might help.
Also I did not get it right probably, if you having issues with growth in some wells of the plate but not in rest? Are you having this problem in wells where you are adding extracts or in your control wells?
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