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T4 phage complementation for Teaching labs


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#1 Mthomas

Mthomas

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Posted 14 January 2011 - 10:42 AM

I am currently working on some labs to be used in a teaching lab. I want to do the old T4 rII complementation test that I remember being part of teaching labs back when I was in grad school.

I purchased some T4 page strains and E. coli K-12 and B from Ward's and Carolina and I've run into a bit of a snag. It is my understanding that the mutant T4 will not form plaques on the K-12(lambda). But I'm seeing small plaques with both the Wild type and the mutant T4. The complmentation experiments have been a total bust as a single mutant phage still produces plaques, so basically I have no assay. I'm wondering if possibly the K-12 is not K-12 lambda or my mutant phage are not mutant. I'm doing some controls on that, and both suppliers have sent me multiple K-12 samples, all with the same result. Tech support at the suppliers, while friendly have no real suggestions. Is Uv exposure a reasonable way to test the K-12 for presence of the lysogenic lambda? I would love to be able to confirm that the K-12 is the right variety as that would rule a huge problem out.

I'm wondering if someone with a bit more experience with this has any suggestions or thoughts. Or some place I can get bonafide K-12(lambda) and not one of the other variants of that strain. I've been working from some papers published in various teaching lab journals regarding titers and OD's to use and have been not been able to contact the authors yet for any suggestions from their side.

Any thoughts or suggestions would be very much appreciated.
Mthomas@ncf.edu




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