1 reply to this topic

[Wera]

Hey,

I have some problems with obtaining PCR product for cloning (in pENTR vector). After trying many times with different conditions and primers etc I used PCR with modified nucleotide - 7-deaza-2'-deoxyguanosine- (which is used for GC rich regions for sequencing aims mainly) and i have proper PCR product for my gene of interest (checked by sequencing).  Now the question is if this 7dcGTP has some influence on translation, because if yes I can't use it in my research. Does anyobody know sth about this?? I haven't found anything about this on the internet...

Best,

Wera

[phage434]

By the time it is cloned, transformed, and transcribed, it will be a plain G, so no, it should be fine.  To double check, you can sequence the plasmid after cloning.  If you are doing in vitro expression directly from the pcr product, then I could imagine you might see an effect.