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.
This kit is intended for use in conjunction with Edvotek Transformation Kits 222, 223 or 303.
For 10 Lab Groups
Set Up (30 min) PCR (2 Hours or Overnight) Electro. (45 min)
Kit Includes: instructions, control BactoBeads™, ampicillin, IPTG, ReadyPour™ Luria Broth Agar (sterile), Petri Plates, sterile loops, PCR EdvoBeads™, control DNA and primers, microcentrifuge tubes, PCR tubes, agarose, DNA ladder, practice gel loading solution, 10x Gel Loading Solution, buffer, FlashBlue™ stain, InstaStain® Ethidium Bromide
All You Need: micropipets to measure between 5 and 50 µl, tips, 37°C incubator, microwave or hot plate, thermal cycler, electrophoresis apparatus, power supply, UV transilluminator.
Storage: Some Components Require Refrigerator and Freezer Storage