Biotin Pcr primers and anealing temperature

Karl Voss karl at hobbes.chem.ualberta.ca
Thu Aug 22 11:01:17 EST 1996


Hi,

Does anyone know if having 5' biotin on pcr primers lowers the anealing 
temperature that they need.  I have been getting very good pcr results with
a non-biotinilated primer set that have short 5' sequence overhangs to be
used for directional cloning.  I wanted to directly sequence the pcrs so 
I got a biotinilated primer made that did not have the extra 5' bases.
The resulting pcr bands are very weak.  I have quantified the primers 
carefully and assume that they are of high purity (they were purchased from 
Research Genetics).

Has anyone run into similar situations?  If so I would like to hear
from you.

Thanks to everybody for reading this.

Karl

--
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If it ain't broke it doesn't have enough
features yet

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Karl Voss
Department of Chemistry
University of Alberta
Edmonton, Alberta, Canada
T6G 2G2

karl.voss at ualberta.ca
phone 403-492-0222
fax   403-492-8231



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