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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