I am trying to write the primers for cloning DCoH httpswwwn

I am trying to write the primers for cloning DCoH ( https://www.ncbi.nlm.nih.gov/nuccore/50086629 ) using vector pET28b ( https://www.staff.ncl.ac.uk/p.dean/pET28.pdf ), and constructing the vector so that DCoH has no N terminus additions and a C-terminal histidine tag.

I am struggling with writing the sequence for the finished PCR product. I know the vector has a C-terminal addition tag, and that I don\'t have to include the his tag sequence in my PCR product because the vector includes one, but I\'m having trouble choosing the resitriction sites and making sure the reading frame is correct when my PCR product is inserted into the vector.

Thanks for the help!

Solution

hi.

you may try with restriction site Bgl 2 and Not 1 for your pcr amplification.