GFP Transformation Extension: Colony PCR (323), Edvotek

Use PCR and gel electrophoresis to analyze bacteria transformed with GFP.

• For ten lab groups
• Set up: 30 minutes PCR: 120 minutes Electrophoresis: 45 minutes
• Intended for use with Edvotek kit 222, 223, or 303

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.

Storage: Some components require refrigerator and freezer storage upon receipt.

Ordering information: DNA electrophoresis, thermal cycler,micropipets: 5 to50 µl, UV or blue light transilluminator, 3 water baths (if performing experiment without a thermal cycler), white light box (optional), microcentrifuge, and microwave or hot plate.

Delivery information: Includes Instructions, PCR EdvoBeads™, colony primer mix concentrate, control DNA concentrate, ampicillin, IPTG, EdvoQuick™ DNA Ladder, TE buffer, bactoBeads™, ReadyPour™ luria broth agar, petri dishes, , ampicillin, IPTG, ReadyPour™ luria broth agar (sterile), UltraSpec-Agarose™, electrophoresis buffer (50X), 10X gel loading solution, InstaStain® ethidium bromide, FlashBlue™ liquid stain, petri plates, sterile loops, microcentrifuge and PCR tubes and wax beads.