pGLO Lab Report Essay

821 WordsApr 24, 20144 Pages
Bacterial Transformation Lab Report Backround: The plasmid pGLO contains an antibiotic-resistance gene, ampR, and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli, so if E. coli, so if E. coli cells contain the ampicillin-resistance gene, the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus, transformed E. coli cells containing ampicillin-resistance plasmids can easily be selected simply growing the bacteria in the presence of ampicillin-only the transformed cells survive. The ara control region regulates GFP expression by the addition of arabinose, so the GFP gene can be turned on and…show more content…
6. Incubate both microcentrifuge tubes on ice for fifteen minutes. 7. Take both tubes out of ice and immediately place in incubator at 42٥C for 90 seconds. 8. After place both tubes back in the ice for two minutes. 9. Add 200uL Luria Recovery Broth to both microcentrifuge tubes. 10. Let both the tubes rest at room temperature for 10 minutes. 11. During the 10 minutes, get the LB agar and LB+AMP agar plates ready. Mark your plates with the transformation tube mixture to use (+ or -), the lab group names, and the date on the top of the dishes. 12. Add 100ul of the pGLO transformation cell mixture to the center of the agar surface of the corresponding LB agar and LB+AMP plates. 13. Use a sterile plastic loop to distribute the cell suspension evenly on the plate by “skating” the loop back and forth across the LB agar plate several times. 14. Use the same loop and technique to spread the same cell suspension (+) on the LB+AMP agar plates. Dispose of the sterile loop in a beaker of germicide. 15. Repeat the procedure by spreading the (-) transformation cell mixture to each of the (-) labeled LB and LB+AMP plates. Be sure to use a fresh plastic loop for the ‘None’ transformation mix. 16. Stack your group’s set of plates on top of one another and tape them together. The plates should be left upright position to allow the cell suspension to be absorbed by the agar. 17. Place the plates in an inverted position (agar side on top) in a 37٥C
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