I am looking to target a specific gene for amplification. In the literature, a group have targeted the same gene, in the same way using a set of primers with a Kozak consensus sequence at the 5' end.
The Kozak consensus sequence is new to me: If I remove it from the primers, will they still work?
Thanks!
Hey!
If it is new study about that (Refer Campbell Biology). Start Codon ATG itself is present in Kozak consensus sequence and play major role in translation initiation. I wonder how do you remove that. It is part of the gene.
It is better to keep the Kozak consensus sequence to ensure high protein expression. PCR primers will still work if you put the Kozak sequence at the 5'-end of primers. The 3'-part of primers should, however, be identical with your gene template since PCR reaction extends from 3'-end of the primer; hence perfect match between 3'-end of primer and template are needed.
They should work. But if you're trying to get a protein expression, I would recommend including the fragment.
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