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Two start codons" the TAG and the gene"


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#1 saad

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Posted 08 July 2011 - 11:57 PM

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
SA

#2 bob1

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Posted 10 July 2011 - 04:39 PM

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.




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