Biotin Pcr primers and anealing temperature

Karl Voss karl at
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.



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

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

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

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