Does CIAP cause ligation problems?

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

Steven Goldberg (goldberg at 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.

* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - -  Methods FAQ list -> - - -  *
* -  TIBS column archive -> - -  *
* - The BEST Molecular Biology HomePage ->  *

More information about the Methods mailing list