HOW TO GET A Sharp fragment using PCR product as template

G.E. Woodward gew23 at HERMES.CAM.AC.UK
Thu Mar 18 16:17:14 EST 1999


Dear Methods,

	I am trying to amplify a PCR product from a cDNA library using a
vector primer (T7) and a degenerate oligonucleotide for the center of the
gene.  I get two products which are faint and want to gene clean them and
re-amplify using the products as a template.  However, when I attempt to
do so using the same primers I tend to get a smear.  Do you have any
recommendations as to how to clean and re-amplify these fragments using
degenerate oligonucleotides in order to get a sharp clean fragment?


						Yours sincerely,


						Geoffrey Woodard





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