bacterial culturing for BAC - (Jun/13/2008 )

Hi - I have a question. I've inoculated 5 mL cultures for 3 days (new culture every day inoculated with the previous culture). Should I be concerned about deletions or mutations in the BAC due to prolonged propagation? I use chloramphenicol as a selection.

Thanks!

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Yes. Definitely. Rearrangement is the horrifying word rightly associated with BACs.

If you are culturing at a lower temperature, rate of recombination would be low, but still there.

It also depends upon the bacterial strain you are using, but as a rule of thumb, always start a culture (not to mention BAC culture) from a single colony on the plate.

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