lysis problems with bacteriophage cultures

Helene Puehringer h.puehringer at iam.boku.ac.at
Fri Oct 18 09:27:12 EST 1996

has anyone an idea why my bacterial cultures (E.coli KW251) inoculated with lambda-GEM11
do not lyse after overnight incubation at 37°C. The small scale liquid cultures of phages
was performed according to Maniatis et al.. 0.1ml bacteria (grown overnight in NZY medium
+ 0.2% maltose) in 10mM MgSO4 were incubated with 0.1ml bacteriophages in SM buffer 
for 15min at 37°C. Then 2ml NZY medium were added and the cultures were grown overnight 
at 37°C. But no lysis occurred!!!!
Neither addition of an equal vol. of medium and further incubation for 3h nor incubation
at elevated temperature (39°C=optimum for lambda-phages to grow) could induce lysis.
What is wrong with my cultures? Is the ratio bacteria:bacteriophages at the initiation of the
culture critical for complete lysis? If yes, how should it be?
Please, let me know your suggestions. I would be grateful for any help!

Thanks, Helene.

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