In this experiment, students will explore the biological process of bacterial transformation using¬†E. coli¬†and plasmid DNA. At the end of the activity, students will have experience observing and analyzing acquired traits (ampicillin resistance and fluorescence) as exhibited by transformed bacterial cells.
‚Ä¢ Explore genetic engineering and the central dogma with this elegant experiment
‚Ä¢ Transform¬†E.coli¬†with pFluoroGreen‚Ñ¢ - a plasmid containing genes for ampicillin resistance and the green fluorescent protein
‚Ä¢ Select for transformed cells using LB-ampicillin plates and calculate transformation efficiency
‚Ä¢ Expose transformed cells to IPTG to demonstrate differential gene expression
‚Ä¢ New transformation protocol has high transformation efficiency = great results!
Instructions, BactoBeads‚Ñ¢ E. coli GFP Host, supercoiled pFluoroGreen‚Ñ¢ plasmid DNA, ampicillin, IPTG, CaCl2, Growth Additive, ReadyPour‚Ñ¢ Luria Broth Agar (sterile), Luria Broth Medium for Recovery (sterile), petri plates (small), petri plates (large), plastic microtipped transfer pipets, wrapped 10 ml pipet (sterile), toothpicks (sterile), inoculating loops (sterile), microcentrifuge tubes
All You Need:
Automatic micropipette (5-50 ¬µl) and tips, two water baths (37¬∫C and 42¬∫C), thermometer, incubation oven (37¬∫C), pipet pumps or bulbs, ice, marking pens, bunsen burner, hot plate or microwave, hot gloves, long wave UV light