I want to clone a PCR fragment into a expression vector I made. The
polylinker has a Pvu II site and need to make a in-frame fusion into this
1) Clone the PCR fragment as blunt by using a polymerase that does not
2) Clone the PCR fragment by first polishing the extra base and blunt clone
3) Add a extra T to the pvu II site either by taq polymerase or terminal
deoxy transferase and clone in a PCR fragment with an A over-hang.
4) Design a restriction site into the PCR primers to clone
Which one of the above is likely to be the easiest? I will summarize the