I do not have a plasmid containing the NLS. I was planning to add the NLS to the sequence of a primer. My question is: Do I just place the sequence straight after the Start codon without a linker sequence?
Hi Julia,
I suggest to use a small linker and overal to try add NLS at both N and C terminus and test wich is the best!!!
Good luck
Cesare
Hi Julia,
I suggest to use a small linker and overal to try add NLS at both N and C terminus and test wich is the best!!!
Good luck
Cesare
My hunch is a linker is probably not required as NLS is not very bulky. But save yourself some time and do all combinations at the same time to save yourself some workload.
As primers are pretty cheap. I would do N and C terminus and with or without linkers. And then test which gives the best expression and localisation.
I also use NLS in my constructs without any linker.I usually have it just at the C-terminus.
Just as an additional comment, in case it does not work. Depending on which type of cells you use, think about which NLS is the best. There are good ones and bad ones.
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