P1 transductions

Dr. Duncan Clark duncan at genesys.demon.co.uk
Tue Jul 9 14:43:50 EST 1996


In article <31E2A6F3.6978 at igmors.u-psud.fr>, David Clarke
<CLARKE at igmors.u-psud.fr> writes
>Hi,
>I know that P1 transductions are supposed to be "easy" but I'm
>hoping that someone can help me out. I cannot (and I don't know
>why) manage to get my titres of P1 sufficiently high for
>efficient marker transfer. Although I can sometimes get good
>metabolic marker transfer (e.g. arabinose) I have yet to be 
>able to transduce (repetitively) drug resistance markers with the
>same lysates. I have been following the protocols in Miller (1992), so
>if anyone has any suggestions I would be very grateful.


Silly as they may be make sure your strains are recA+. I had real
problems until this was pointed out to me! To get really high titres we
landed up on some strains just growing up 100ml and PEG ppting the
pahge. Crude but it worked. We have also used the Cm P1 lysogen. This
worked nicely for us. We never got high frequencies of transductions but
we did get what we wanted. We only did it to make a couple of strains so
never really played with the system to get it working at better
efficiency. Miller's CSH handbook was useful as was some advice from NEB
and this newsgroup.

Duncan  
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My mind's made up. Don't confuse me with the facts!
Duncan Clark
GeneSys Ltd.




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