PCR using a primer with a dUTP

Julie Johnson julie at DNA.CHM.JHU.EDU
Mon Jul 15 16:17:10 EST 1996


I am currently using PCR to make a 189 bp DNA fragment. All of the
following oligos discussed are deoxyoligonucleotides.  One of my oligos
(#1) is a "normal" 32 bp oligo and it works just fine with my second
oligo (#2) in the same reaction.  Now the problem, I have another oligo,
let's call it #3, that is the same as #1 except 5 bases from the 3' end
of the oligo there is a dUTP instead of a dTTP.  The PCR reaction
conditions are the same whether I use #1 and #2 or #3 and #2. For some
reason when I use #3 I get *no* PCR product.  Anyone have any experience
with PCR with dUTP in the primers?  Any suggestions? Meanwhile, back at
the ranch, I'll be trouble shooting.  Joy. 
-- 
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Julie Johnson		   _~0    julie at dna.chm.jhu.edu	
The Johns Hopkins Univ.  _`\<,_   http://dna.chm.jhu.edu/~julie	
Dept. of Chemistry      (_)% (_)  (410)516-7472
Baltimore, MD 21218		  (410)516-8420 FAX



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