classic cloning with restriction enzymes into empty pcDNA3.1D - (May/17/2013 )
I have a construct that I want to insert into pcDNA3.1D, but I want to do it with regular cloning using restriction enzymes and not TA cloning using the TOPO sites. Whilst discussing the synthesis of my construct, one of the companies I had contacted did not recommend restriction site cloning my construct into an empty pcDNA3.1D vector because they have no experience with this and they are not sure it would work. I don't see why it wouldn't work. Does anyone have any experience with cloning using an empty pcDNA3.1D vector? Are their concerns justified?
Thanks a lot!
It should work fine, so long as there are compatible sites, I've cloned into pcDNA3.1 a few times using regular cloning, with no problems. From your post I take it that you are getting a gene synthesized and then going to subclone that - which should be fine.