Dear Hadi,

Certain palsmid already gives you the option of C-terminal FLAG tag. You have to clone your insert in-frame of that tag. if you donot have access to such plasmid add codons of FLAG tag followed by a stop codon to 5'-end of your reverse primer. Once again check the frame before ordering primers

Dear Junaid

Thanks for your answer, but I am going to clone my insert into a plasmid which does not give this option

Hadi

If you donot have access to such plasmid add codons of FLAG tag followed by a stop codon to 5'-end of your reverse primer. Once again check the frame before ordering primers

Dear Junaid

In this case the target gene will translate but the flag tag will not translate as a result the protein will not be tagged

thanks

No! see below the C-terminus of translated protein should be like it

XXXXFLAG.

here xxx is your protein followed by FLAF and stop codon (.)