Can someone please help me with Gibson Assembly of my ~4KB insert into a pAAV vector.
I have followed all the steps but having problems.
Help with primer design, reaction conditions etc and if anyone could look at my specific sequences would be really helpful. I can send sequences etc if anyone is willing to look.
Prior to the PCR reactions, do the vectors containing the insert or the destination vector need to be linearised? I did not think that was necessary so did not do that.
Any help would be greatly appreciated.