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Two start codons" the TAG and the gene" - (Jul/09/2011 )

Hi Everyone
I've read that when you clone your gene post N-terminus tag(His or GST)you might see two bands because of the existence of another AUG in the gene.
I was wondering when I want to clone my gene or part of it after Tag, is it necessary to add "AUG" start codon in the Fwd primer or I shouldn't add this start codon. What the right way to clone construct after tag.

Thanks heaps


You can do either, both work absolutely fine. If you add a kozak sequence before the "tag" start codon, this will help recognise the correct one.