This project involves transforming bacteria with a GFP plasmid to study genetic engineering. Students will measure transformation efficiency by counting fluorescent colonies.
If bacteria are successfully transformed with a GFP plasmid, then the colonies will fluoresce under UV light.
As an Amazon Associate, this site earns from qualifying purchases.
Prepare LB agar plates with antibiotic.
Mix E. coli culture with calcium chloride solution.
Add GFP plasmid to the bacterial suspension.
Incubate on ice for 30 minutes.
Heat shock the mixture at 42°C for 90 seconds.
Return to ice for 2 minutes.
Add recovery broth and incubate at 37°C for 1 hour.
Spread bacteria onto LB agar plates.
Incubate plates overnight at 37°C.
Use UV light to identify fluorescent colonies.
Count the number of fluorescent and non-fluorescent colonies.
Calculate transformation efficiency.
Successfully transformed bacteria will form fluorescent colonies under UV light. The transformation efficiency will depend on the proportion of fluorescent colonies to total colonies.
Plasmid presence
Number of fluorescent colonies
Bacterial strain, Incubation time, Temperature, Nutrient media
Handle all bacterial cultures using sterile techniques and dispose of waste properly.
Include images of fluorescent colonies.
Explain the significance of GFP in biotechnology.
Discuss factors affecting transformation efficiency.
A GFP plasmid carries the gene for Green Fluorescent Protein, used as a marker in genetic engineering.
Calcium chloride increases the permeability of bacterial cell membranes for DNA uptake.
It is the ratio of fluorescent colonies to total colonies expressed as a percentage.
