Does CIAP cause ligation problems?

Paul N Hengen pnh at ncifcrf.gov
Fri Jan 10 15:42:59 EST 1997


Steven Goldberg (goldberg at bms.com) wrote:

> For the last two months I am trying to perform a non-directed ligation. 
> Therefore I tread my vector with CIAP (Promega). However, till now I wasn't 
> successful.

: I gave up on CIAP a long time ago because of the difficulties I had
: getting fragments to ligate.  I think there is a lot more exonuclease
: activity associated with this enzyme than manufacturers are willing to
: admit.  Anyway, the best alternative for routine subcloning or plasmid
: library construction has been pZeroI by Invitrogen, where I routinely get
: 90% recombinant plasmids.
[snip %<----------]

I had the same problem with CIAP recently. I was trying to construct a
phage display library and had to give up on it because I had an enormous
background of empty books in the library :-(  Now, if only someone could
construct a pZero phage display vector with ccdB in it -> THAT would sell
like hotcakes! Okay, so I'm giving away this great idea...but I have yet
to be offered a job by a company willing to pay me for making it ;-)
BTW, I'll be leaving here soon because my time is up....Somebody please
offer me a new job :-) I'm willing to relocate to the west coast.

--
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* Paul N. Hengen, Ph.D.                           /--------------------------/*
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