WOW Biolab: Bacterial Transformation

PROBLEM

In this investigation, you will use the process of bacterial transformation to add a new gene to E. coli cells. The new gene will make the bacterial cells resistant to an antibiotic that would normally prevent the bacteria from growing.

MATERIALS

• E.coli Bacteria
• tubes of Calcium Chloride (CaCl2)
• E.coli Plasmids with ampR
• sterile pipette
• ice water bath
• warm water bath
• agar infused with ampicillin
• sterile inoculation loops
• biohazard container
• incubator
• sterile toothpicks

PROCEDURE

1. Drag a toothpick to E. coli petri dish to get a sample of E. coli, then drop the sample into the calcium chloride tube 1.
2. Drag another toothpick to E. coli petri dish to get a sample of E. coli, then drop the sample into calcium chloride tube 2.
3. Use the sterile pipette to move E. coli plasmids with ampR into tube 1.
4. Drop tube 2 and tube 1 into the ice bath for 15 minutes.
5. Move the tubes from the ice bath into the warm bath for 90 seconds.
6. Drop a sterile inoculating loop into tube 1 to get a sample, and spread it in petri dish one with Agar.
7. Drop a sterile inoculating loop into tube 1 to get a sample, and spread in in petri dish 2 with agar + ampR.
8. Drop a sterile inoculating loop into tube 2 to get a sample, and spread it in petri dish 3 with agar.
9. Drop a sterile inoculating loop into tube 2 to get a sample, and spread it in petri dish 4 with agar + ampR.
10. Place all petri dishes in the 37 degrees celsius incubator for 24 hours.
11. After 24 hours, remove the petri dishes from the incubator and observe results.

RESULTS

• Petri dish 1: There were experiences of growth of bacteria, thus, bacteria within the agar and E. coli with ampicillin are antibiotic resistant.
• Petri dish 2: Like Petri Dish 1, bacteria grew, meaning the bacteria within agar with ampicillin and E. coli with ampicillin are antibiotic resistant.
• Petri dish 3: There were also experiences of growth of the bacteria, signifying that the bacteria within the agar and E. coli without the ampicillin are antibiotic resistant.
• Petri dish 4: No bacteria grew, therefore, the agar with ampicillin and E. coli without ampicillin are not antibiotic resistant.