Hi,

I want to generate a HA-tagged fusion protein (HA-tag at protein C-terminus) during transient expression in plants. I'm choosing a PCR method over restriction digestion. Strategy is, a regular forward primer (20 bp length) which can binds from the 5' end of my ORF and a RP with 3' end of ORF (reverse complementary orientation) without stop codon (approximately 20 bp) followed by HA-tag sequence (27 bp), a restriction site (6 bp) and stop codon (3 bp). Reverse primer size will have the length of 56 bp. Will this strategy produce a successful fusion protein?

Thank you