Transducing rec+ to recA1 E.coli with P1

Duncan Clark Duncan at genesys.demon.co.uk
Fri Feb 10 12:26:16 EST 1995


Hi Folks,

We are having serious problems trying to transduce a recA1 E.coli 
(Invitrogen TOP10) to recA+ using P1 lysates of JC10241 (from 
B.Bachmann @ CGSC). JC10241 has Tn10 (TetR) in srl-300 right next 
to recA. The bug is definitely tetR but it just refuses to go across 
to TOP10. This is the standard strain for transducing recA+. The 
lysates appear to be OK, I even did a 250ml lysate and concentrated 
it with PEG/NaCl etc to 1ml. Still no go. We can transduce other 
markers from different E.coli P1 lysates into TOP10 although they do 
go much better into say RR1. I particularly want to use TOP10 'cos it 
delted for lacz and mcr restriction systems. 

All idea appreciated even more so if you have transduced recA+ and 
have a lysate!

Many thanks

Duncan

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