Biotin Pcr primers and anealing temperature

Karl Voss karl at hobbes.chem.ualberta.ca
Tue Aug 27 12:19:10 EST 1996


Hi gang.
 
Just to followup about this biotin thing I though I would let you know what
else I have seen.  I have both biotinilated and regular primers for both ends
of the pcr I am doing.  The primer at one end has a long 5' overhang for 
doing dye primer sequencing with abi M13-21 primer sets.  When I use the 
new biotinilated primer containing the long overhang with the non-biotinilated
primer on the other end my pcr works fine with no changes in the melting 
temp.  When I use the biotinilated primer without the overhang and the regular
primer with the overhang is when the annealing temp drops 3 degrees.  

Although I can't say from these limited experiments it appears that the 
biotin disrupts base pairing of the base(s) nearest to it.  If those bases
arn't involved in annealing (as when a 5' overhang is present) then the
biotin doesn't have an effect on the pcr.

Good luck to all.  Thanks to everyone for helping to work this problem
out so quickly.

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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