Epitope tagging - for the lazy bones - (Nov/26/2007 )

I wish to add an epitope tag at the C-terminus of my gene of interest (GOI). I have already made a GFP fusion construct and there is one unique restriction site right in the middle of the GOI and the GFP coding region. I'm wondering if I can design and order a synthesized double stranded DNA containing v5 or myc tag + the stop codon + the appropriate overhangs and then just ligate it into the original construct after excising the GFP region. I hope this was easy enough to follow. Thanks for any feedback.

-deepbluecheese-

It should work as long as the restriction site is not a weird one.

-scolix-

The unique site I'm targeting is spe1.
For making the double stranded DNA (containing the tag, RE site etc) I can just mix a set of fwd and rev. primers right?

-deepbluecheese-

QUOTE (deepbluecheese @ Nov 27 2007, 12:48 PM)

The unique site I'm targeting is spe1.
For making the double stranded DNA (containing the tag, RE site etc) I can just mix a set of fwd and rev. primers right?

It should work. remember to add a few extra bases at the end of primers for proper digestion of PCR product.

-scolix-