PCR Subcloning with restriction sites

Tony Schountz tschount at mesastate.edu
Thu Mar 30 19:03:40 EST 2000

If I designed my primers with the first 6 nucleotides for restriction sites, then 20
nucleotides that are gene specific, then the result after PCR (Taq) should have
amplicons with the restriction sites at the very end with an A overhang on the 3'
ends. When this was cloned into pGEM-T, I expected that the restriction sites would
have been preserved. Perhaps I should have the plasmid sequenced to see what
happened to the restriction site?

Thanks again,


Tony Schountz, Ph.D.
Department of Biological Sciences
Mesa State College
mailto:tschount at mesastate.edu

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