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DNA transformation of escherichia coli using plasmids

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Boston Medical Center

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  1. Abstract
  2. Introduction
  3. Methods
    1. Observing colonies of E. Coli
  4. Results
  5. Discussion
  6. Concluding points
  7. References

During this lab, we performed a transformation of genes from one organism to another, more specifically, added genes to a bacteria. Our results displayed much growth (100+ colonies of small size that did not glow) on the +pGLO LB/amp plate. The +pGLO LB/amp/ara plate experienced growth (100+ colonies of mid to small size that glows). The ?pGLO LB/amp plate experiences no growth, while the ?pGLO LB plate experience lawn growth and does not glow. These results are interesting, as they are exactly the results that we expected, agreeing with my hypothesis that the plate containing the GFB gene and arabinose sugar would be transformed.

[...] We then transferred 250 ul of transformation solution (CaCl2) into each tube, and then put both on ice. A single colony of bacteria was then taken from the starter plate by the use of a sterile loop and placed into both + and labeled tubes. The colony was spun until no chunks appeared. We then observed this solution under a UV lamp. Next, we placed a new loop into the pGLO plasmid DNA stock tube. This loopful was then mixed into the +pGLO solution, while the ?pGLO solution was left alone. [...]

[...] This was followed by us pipetting 100 ul of the transformation and control suspensions onto the correct nutrient afar plate on the + plate, - on the plate). We then spread this evenly over the surface of the plate. We finally handed out plates over to our TF, who incubated then at 37 degrees Celsius for 24 hours. RESULTS Table 1 provides the results that were yielded from this experiment: Plate: Growth of colonies? Glow under ultraviolet light? +pGLO LB/amp Many shiny colonies No Medium to very small sizes, grouped with like sizes. [...]

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