Lambda libraries

afc at gnv.ifas.ufl.edu afc at gnv.ifas.ufl.edu
Tue Oct 13 10:30:16 EST 1992


In article <1992Oct13.100236.90319 at vaxc.cc.monash.edu.au>, mic255z at vaxc.cc.monash.edu.au writes:
> Dear Netters,
> 
> I am interested in using a lambda system to do some primary cloning.
> Does anyone have any thoughts on what system is better ie., what are the
> relative merits of lambda-ZAP and lambda-DASH compared with EMBL, GEM
> or gt11 apart from obvious things like unique cloning sites.

The ZAP sorts of vectors are great, since they make subcloning simple.
We have been using Promega's lambda-GEM vectors, and instead of fooling
around trying to find out how to subclone six different fragments from
each clone, we get the subclones the next day.  (This is a plug for the
general approach, not Promega in particular.)

> Primarily, I want to clone bacterial outer membrane proteins using specific
> antisera to detect recombinants, but I also want to generate lambda libraries
> to store for future (unknown) use.  Does this later the comments generated 
> by the above question?

In the dark ages of the 1970s library construction was black magic.  We
constructed libraries and replicated a successful one for years.  It
was worthwhile to plan construction to maximize the percentage of the
genome represented and to try to get as big a library as possible.  Now
that you can buy fancy vectors, packaging mix that works EVERY TIME!,
and good hosts it is really pretty easy to make a library.  Even cDNA
libraries are not that hard.  It is no longer useful to try to construct
the "perfect" library.

Figure out what fragments you want to clone, configure your library 
construction *to those fragments*, and throw it away afterwards.  You
will be ahead in the long run.  IMHO, of course.

Andrew Cockburn

> ANd finally, lambda-ZAP seems a very good system and I am leaning towards
> this.  Has anyone used this system with less than satisfactory results?
> 
> Thanks in advance,
> Helen Jost,
> Department of Microbiology,
> Monash University.



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