Dear colleagues,
I am synthesising some RNA which contains the IRES, ORF, 5' UTR and 3'UTR.The original DNA ORF will be replaced with a new ORF with Gibson reaction. Although, there will be a 'scar' of one nucleotide between the end of the new ORF and the 3'UTR.
Do you think it would matter, and the Gibson primers should be designed to skip this nucleotide, or it doesn't matter? Thanks!
Kind regards,
Maria