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One Step Growth Curve

| posted 14 Jan, 2019 16:42
We are trying to calculate burst size for our phage but are having trouble getting a one step growth curve to work reliably. Is there a popular protocol that people use with smeg that works well? Thanks!
| posted 14 Jan, 2019 16:48
One step growth curves in Mycobacteria are really hard. I've tried a number of times, but never really been satisfied with the data; it just doesn't look as nice as ones with E. coli or Salmonella phages.

In general, we try for exponential cells with an OD of 0.4, and MOI of at least ten. We do the infection in a small volume (so you pellet your cells, resuspend in 1/10 or even smaller a volume); add phage; wait 15 min; and then bring back up to the initial volume. Shake at 37 for 3 hours or so while you sample.

Latent periods seem to run around 3 hours-ish.

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