Question

Lab: P1-RFP Plasmid Extraction

In the lab, the goal is to assess if you did indeed extract P1 plasmid containing RFP vs. plasmid containing only P1.

What is the purpose of the loading a P1 plasmid alongside the P1-RFP plasmid? What banding pattern (sizes) do you expect from each sample when looking at agarose gel?

Answer 1

Loading a P1 plasmid alongside the P1-RFP plasmid helps to determine whether the RFP is inserted in the P1 plasmid properly or not. It is used as a negative control.

When looking at agarose gel, we would expect a larger banding pattern (sizes) for P1-RFP plasmid in compare to P1 plasmid. The reason behind that in case of P1-RFP plasmid, RFP is inserted in P1 plasmid while P1 plasmid is only the plasmid, there is no insertion. Therefore, in agarose gel, P1 plasmid will run faster than P1-RFP plasmid, and cover a greater distance.