2 replies to this topic

[Brian Jackson]

I am trying to design a forward primer to amplify an ORF to make an mRNA to express a protein without the fusion tags in a bacterial system.  So the vector goes something like this

[Promoters][Ribosome binding sequence][fusion tags (V5/His x6)][Stop codon]*[Ribosome binding sequence][9-10 nucleotides][ATG-my protein][Stop codon].

My question is: Do I need to add any spacer nucleotides between the first stop codon and the next ribosome binding sequence (at the *), or can they be right next to each other?

[pDNA]

can be right after each other ...e.g. see the lac operon!

so no special spacer needed!

Regards,
p

[Brian Jackson]

Great, thanks!