HELP: ligation (used to be Re: cip & geneclean)

Stuart Brown browns at ccu.umanitoba.ca
Fri Aug 21 10:17:01 EST 1992


>>> 
>>>We have been attempting to clone a fragment of DNA into a high
>>>copy-number vector (a derivative of the pUC plasmids).  We have had some
>>>problems with incomplete digestion of the vector causing high
>>                ^^^^^^^^^^^^^^^^^^^^
>>>background, thus making identification of recombinant clones difficult
>>>(using lacZ blue/white identification).  
>> 
>Thank you (in advance) for any assistance with our problem,
>Vernon Coyne
>coyne at uctvax.uct.ac.za
>
>  

A simple method for reducing background from uncut vector: after digestion and
dephosphorylation, run your cut vector on a gel and cut out the band of linear
molecules.  Ligate in agarose or clean DNA out of agarose with your favorite
method.

	-Stu

-- 
Stuart M. Brown                             If you can remain cool when all 
U. of Manitoba, Dept. Plant Science         Around you are in panic,
Winnipeg, Manitoba, CANADA
browns at ccu.umanitoba.ca            Then you surely misunderstand the situation



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