Why is it important to have an antibioticresistance gene on

Why is it important to have an antibiotic-resistance gene on the vector? After the transformation, we allow cells to incubate at 37oC for 45 minutes before plating them onto an LB + kanamycin plate. Why did we do this? What will happen if we don’t?

Solution

The antibiotic resistance gene allows for selection in bacteria. This is very important to allow cells to incubate at 37⁰C for 45 minutes afterthe transformation both the E.coli to recover from the heat shock, and to allow them to start to express the antibiotic resistance genes (ampicillin). This should not be too much longer than 45mins as you do not want them to divide, but just recover.