lysis problems with bacteriophage cultures

Artur da Camara Machado iam at mail.boku.ac.at
Thu Oct 17 03:48:48 EST 1996


Hallo,
has anyone an idea why bacterial cultures (KW251) inoculated with lambda-GEM11
do not lyse after overnight incubation at 37°C. The small scale liquid culture of phages
was performed according to Maniatis et al.  0.1 ml bacteria (grown overnight in NZY
medium + 0.2% maltose) in 10mM MgSO4 were incubated with 0.1 ml bacteriophage
suspension in SM buffer for 15min at 37°C. Then 2ml prewarmed NZY medium were 
added and the culture was 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?
I would be grateful for any help. Please, let me know your suggestions!

Thanks, Helene.



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