subcloning

Richard Daines rdaines at pcix.com
Fri Jun 7 09:40:10 EST 1996


Michael Witcher wrote:
> 
>  I am trying to subclone a 2900 bp insert into a bluescript vector,also ~2900 bp in length, and it's not working. The main problem is that the 
dephosphorylatio
> w
> w:wq
> :wq
> wI've been suvcloning 5000bp fragments into Invitrogen's pZer0-2 kan vector.  
It's a "zero" background vector that requires no alk phos treatment so, if 
necessary, you can treat your fragment to cut down on multiple inserts.  
Anyway, could you do a two step construction by first cloning into pZer0 and 
then reisolating a fragment from the vector using pZero's restriction sites 
which may be more compatible with your Bluescript.
-- 
Richard J. Daines
Mystic, CT

rdaines at pcix.com



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