Cloning large fragments

joe kramer kramer at
Thu Sep 2 10:35:07 EST 1993

Hello all;

   I need to make a multistep construction that approaches 
30 kb.  I was wondering what hints,  help, and warnings
y'all might have concerning such an undertaking.  Some
of the questions I have are as follows:  

Are pUC based vectors somewhat limited in their 
ability to carry large segments...Is pBR better
or is there something else.

Are certain bacterial strains more or less disposed 
for the task?

Are there any tricks and rules-of-thumb concerning 
ligations involving large inserts (20kb?)

Joe Kramer

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