cloning problems

theodorn at medlib.georgetown.edu theodorn at medlib.georgetown.edu
Wed Nov 11 10:58:23 EST 1998


In article <72a5bi$k3m at panix3.panix.com>,
  iayork at panix.com (Ian A. York) wrote:
> In article <36488C28.9E6309D1 at gengenp.rug.ac.be>,
> Esbjorn Fiers  <esfie at gengenp.rug.ac.be> wrote:
> >Thanks for your reaction. I should have mentioned that it is quite a
> >trickysubcloning, I think. It is with 3 fragments. That is one of the reasons
>
> My opinion is that it's rarely a timesaver to do a three-fragment cloning.
> You'd usually get your result faster with a series of subclones, each with
> two fragments.  Run your fragments through different plasmids if
> necessary to get the appropriate fragments.
>
> Ian
> --
>     Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
>     "-but as he was a York, I am rather inclined to suppose him a
>      very respectable Man." -Jane Austen, The History of England
>

I'm with Ian here.  With three fragment cloning you often have to start over
anyway, so you might as well do it in two steps. Sometimes the
"intermediates" are useful anyway (making probes, e.g.). You can use
minipreps when constructing subclones, so it doesn't take that much extra
time.

Nick Theodorakis

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