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Suspicion of phage infection?


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#1 spn25

spn25

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Posted 02 October 2004 - 03:19 AM

Our lab has recently suffered from protein expression problem..

We manage to get the overnite cultures(5ml of 2xYT plus a single colony plus 5 ul of ampicilin) inoculated and the products seemed to be coludy enough.
However, when we try to inoculate the overnite into 1 L of media, the media first turn cloudy, OD reaches 0.2 or less, and then decreases, with visible clumps of cell debris precipitated. As a result, we could not carry out subsequent induction at all.
The most annoying part is that this happens on and off, for example, prearing 10 flasks, 8 were affrected by the problem and 2 were OK...
This renders further pain and puzzle in trobleshooting the issue
We tried to clean the bug room with virkon and the problem still exists.....
Could anyone give us some advice?

Edited by spn25, 02 October 2004 - 03:44 AM.





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