CIPPers for breakfast

Pendragon lab_winoto at maillink.berkeley.edu.
Wed Aug 3 21:53:38 EST 1994


In article <30orik$qj1 at mserv1.dl.ac.uk>
"Matthias Zeiner" <mzei at sun0.urz.uni-heidelberg.de> writes:

> Use less CIP. You need as little as 0.01U / pmol DNA ends. Also, you should 
> run the reaction in an appropriate buffer. To my knowledge CIP requires 
> Zn++ Ions, which are not contained in the Boehringer H buffer.

The NEB buffers also don't contain Zn++ ions but they work just fine
for NEB cip.  I've made panels of plasmids cut with various polylinker
RNz followed by NEB cip (5 units/10 ug DNA) and its worked well.  My
guess is that the cleanup phase of either the vector or the insert is
to blame here.

Pendragon



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