Problem:

Cloning, Transformation Ligation

Question 1: DNA ligase needs to have a source of energy in order to create a new phosphodiester bond between different DNA fragments. It also uses Mg+2 as a cofactor. What compounds might you expect to find in the buffer we added to the reaction?

Question 2: What would happen if the plasmid we used lacked any antibiotic resistance marker? How could we tell if a bacteria was transformed with this plasmid?

Provide a brief explanation for your answer.