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Cloning problems

Mike Sussman sussman at cvm.msu.edu
Fri Sep 13 09:17:49 EST 1996


In article <519cv0$qhi at net.bio.net> Miss Golden Clone <zxm at pasteur.fr> writes:
>From: Miss Golden Clone <zxm at pasteur.fr>
>Subject: Cloning problems
>Date: 12 Sep 1996 09:15:28 -0700

>Hi everybody,

>today I have  a really serious question t everyone beeing familiar with
>molecular biology.
>Here my problem:

>Since more than a century now, I am trying to clone a PCR fragment, which
>I designed XhoI-XhoI, into an Xho-NheI. However, since today I have not
>been able to get a single positive clone. Why?
>Does anyone has valuable suggestions, I really would appreciate it.
>Thanks in advance.

My friend, since I was a young molecular biologist in New Orleans, well over a
decade ago, I have used the free offering of molecular biological suppliers to
determine compatible cloning sites.  These companies provide tables of what
cleavage sites can be religated to others with high efficiency. Upon reading
your question I randomly grabbed a catalog, turned to the appendix and lo and
behold found the answer.  XhoI and NheI are not compatible.  You are no doubt
a valuable customer to your MolBio Company rep there in France, so I am sure
you can get a catalog easily and for free. In the meantime try XhoI and SalI
or NheI and XbaI.  Good luck in the future and happy cloning.
Mike





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