Long PCR primers

Andrew J. Doherty a.doherty at bristol.ac.uk
Mon Oct 14 05:49:47 EST 1996


HI every one

I've got a PCR primer which is 76 nucleotides long, containing the c-myc
epitope tag and three overlapping restriction sites but only 12
nucleotides at the 3' end are complementary to the sequence I'm trying
to amplify. Is this a feasible primer to use? I haven't had it
synthesised yet, and just wanted some feedback on whether I was barking
mad to even attempt to PCR with such a primer. Will I have to start at a
lower annealing temp for a few cycles or would a single temp PCR do it?
Just for the record, I'm amplifiying GFP from a plasmid to make fusion
proteins.

Ta very much

Andy
-- 
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   Dr Andrew Doherty                          
   Department of Anatomy                   
   School of Medical Sciences             
   University Walk                               
   Bristol                                            
   UK
   BS8 1TD

   e-mail  A.Doherty at Bristol.ac.uk

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