GFP Transformation Extension: Colony PCR

Colony PCR represents a simple and easy way to determine whether cloning and transformation experiments were successful. In this experiment, students will use colony PCR to analyze bacteria transformed with pFluoroGreen. A single colony will be used as the DNA template for PCR. The resulting PCR sample will then be analyzed using agarose gel electrophoresis. If the bacteria have been transformed successfully, a PCR product representing the GFP gene will be produced. A bacterial housekeeping gene is amplified at the same time as a positive control. The presence of both bands is indicative of a successful transformation experiment.