PCR

Martin Offterdinger a8803349.nospam at unet.univie.ac.at
Fri Jul 10 03:11:51 EST 1998


On Thu, 09 Jul 1998 08:46:05 +0200, carin <carin.jarl at plantcell.lu.se>
wrote:

>>Hi there,
>>We want to put on sites for restriction enzymes on the ends of our gene
>
>>of interest by PCR. Would anybody have a manual, a reference  or tips
>on
>>what to consider etc?
>>
>>Thanks,
>>Carin Jarl
>>Plant Cell Biology
>>Lund University,
>>Lund, Sweden
>>carin.jarl at plantcell.lu.se
>
Carin!
Just add the sequences of the restriction enzyme to the primers as an
overhang. Be very careful in the choice of your enzyme; many enzymes
require additional bases, because they are not able to cut at the very
end of a DNA double strand. A good summary of these topic can be found
in the catalogue of New England Biolabs (NEB).
Hope this is of some help.
Martin
Martin Offterdinger
Internal Med.I,Dept. Oncology
University of Vienna
Austria
E-Mail:a8803349.nospam at unet.univie.ac.at
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