apbio transformation lab

+pGLO LB/amp/ara

Approximately 320 colonies Bacteria spread across plate Bacteria was a pale yellow color GLOW

+pGLO LB/amp

Approximately 320 colonies Bacteria was spread across plate Bacteria was a pale yellow color No glow

1. On which of the plates would you expect to find bacteria most like the original non-transformed E. coli colonies you initially observed? Explain your predictions.

On the -pGLO LB plate you would expect to find bacteria most like the original non-transformed E. coli. This is because everything is allowed to grow inside this plate.

-pGLO LB

Overwhelming amount of colonies, lawn of bacteria Bacteria completely covered plate Bacteria was a pale yellow color No glow

3. Which plates should be compared to determine if any genetic transformation has occurred? Why?

The +pGLO LB/amp and the -pGLO LB/amp should be compared to determine if any genetic transformation has occurred. This is because the +pGLO was given the plasmid which gives it resistance to the antibotic in its plate, while the -pGLO was not. So therefore you can prove that a genetic transformation occurred because the bacteria in the +pGLO LB/amp plate would survive.

amp

ampicillin resistance gene

GFP

Green flourescent protein gene

2. If there are any genetically transformed bacterial cells, on which plate(s) would they most likely be located? Explain your predictions.

If there are any genetically transformed bacterial cells they would be located on the +pGLO LB/amp and the +pGLO LB/amp/ara plates. This is because both these plates have the plasmid inside them which allows for the transformation.

4. What is meant by a control plate? What purpose does a control serve?

In this lab the control plate is a plate that has the same bacteria, same amount, and same temperature as all the other plates. In addition, nothing is done to the control plate that will change the bacteris. In this lab our control was the -pGLO LB plate. The purpose of a control is simple. It is used to compare to the other plates where changes have occurred.

LB

Lysogeny broth, a nutritionally rich medium, is primarily used for the growth of bacteria.

2. Of the E. coli traits you originally noted, which seem now to be significantly different after performing the transformation procedure? List those traits below and describe the changes that you observed.

The amount in each plate and the ability to glow were the only traits that were significantly different after performing the transformation procedure. Among the four plates the -pGLO LB had a significant amount of bacteria in it compared to the three other plates after they all performed transformation. Also after transformation the +pGLO LB/amp/ara plate glowed!

1. Which of the traits that you originally observed for E. coli did not seem to become altered? In the space below list these untransformed traits and how you arrived at this analysis for each trait listed.

The traits that I originally observed that didn't seem to become altered were the color and shape of the bacteria. I arrived at the analysis for each trait by comparing the -pGLO plate with the remaining three plates.

4. From the results that you obtained, how could you prove that the changes that occurred were due to the procedure that you performed?

You could prove that the changes that occurred were due to the procedure that you performed by comparing the plates to the control plate. This clearly shows that only the bacteria that were given the plasmid were able to survive the antibiotic.

3. If the genetically transformed cells have acquired the ability to live in the presence of the antibiotic ampicillin, then what might be inferred about the other genes on the plasmid that you used in your transformation procedure?

You may infer that the other genes on the plasmid are resistant to the antibiotic ampicillin.

-pGLO LB/amp

Zero colonies No bacteria spread across plate

ara

arabinose (part of the e.coli)