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pGLO Transformation Online Lab answers BTEC 1000

Answers to the pGLO Transformation Online Lab for BTEC 1000 at salt la...
Course

Biotech: Engineering Life (IG) (BTEC 1000)

6 Documents
Students shared 6 documents in this course
Academic year: 2019/2020
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Salt Lake Community College

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  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.
    • I’d expect to find the non-transformed bacteria on the pGLO+ LB/AMP/ARA dish, without the introduction of arabinose. I think this because the bacterias would still be intact due to it not being killed by the ampicillin, and therefore look still like the control plate, just without the glow.
  2. If there are any genetically transformed bacterial cells, on which plate(s) would they most likely be located? Explain your predictions.
    • They’d be on the +pGLO plate, because the cells would survive long enough to be transformed, because the transformation gives them the ability to survive the ampicillin.
  3. Which plates should be compared to determine if any genetic transformation has occurred? Why?
    • I think the +pGLO plates should be compared against each other (one with arabinose, one without). I say this because then it is a fair comparison, because they both have the pGLO DNA.
  4. What is meant by a control plate? What purpose does a control serve?
    • A control plate exists to show a further difference in the final product of the experiment, a meaning of comparison between different variables added. In this case, the difference between arabinose and no arabinose, or ampicillin versus no ampicillin.

PAGE 43

  2. How much bacterial growth do you see on each plate, relatively speaking?

    • On the +pGLO plate without arabinose, there’s some bacterial growth, but less dense growth compared to the -pGLO plate with no ampcillin (presumably just e coli and the broth?). On the +pGLO plate with arabinose, there’s more glowing bacterial growth it

looks like, but the spots are smaller. On the -pGLO plate with ampicillin added, there is no bacterial growth. 3. What color are the bacteria?

  • When looking at the +pGLO plate with arabinose added, the e coli is glowing green. The other bacterias shown on the plates are just a greyish color.
  1. How many bacterial colonies are on each plate (count the spots you see).
  • On the -pGLO plate with no ampcillin, there is one very large spot and two small spots. On the -pGLO plate with ampicillin, there is nothing. On the +pGLO plate without arabinose, there’s about 50 spots. On the +pGLO plate with arabinose, there’s like 100+ spots.
  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.
  • One unaltered trait is the shape of the grown e coli that was untouched on the broth plate (-pGLO). You can assume although all of the e coli on the next plate died due to the ampicillin, the shape did not change.

  1. 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.

    • One trait that is very different from the control “blob” on the -pGLO plate with broth is the +pGLO plate with broth and ampicillin. There are colonies now, not a blob, because the majority of the “blob” died when the ampicillin was added, and the colonies that are left represent the e coli's who were able to transform and gain immunity to the ampicillin. The untransformed -pGLO plate with ampicillin shows that they all died.

  2. 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?

    • The other genes on the transformed plasmid have presumably also acquired the ability to live amongst the ampicillin.

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

    • Because e coli doesn’t naturally glow on its own, and did not glow without the presence of arabinose.

  4. Recall what you observed when you shined the UV light onto a sample of original pGLO plasmid DNA and describe your observations.

    • Because the DNA has the pGLO gene in it, the UV light would make it glow green, but ONLY if arabinose is there too.

  5. Which of the two possible sources of the fluorescence can now be eliminated?

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pGLO Transformation Online Lab answers BTEC 1000

Course: Biotech: Engineering Life (IG) (BTEC 1000)

6 Documents
Students shared 6 documents in this course

University: Salt Lake Community College

Was this document helpful?
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.
- I’d expect to find the non-transformed bacteria on the pGLO+ LB/AMP/ARA dish, without
the introduction of arabinose. I think this because the bacterias would still be intact due
to it not being killed by the ampicillin, and therefore look still like the control plate, just
without the glow.
2. If there are any genetically transformed bacterial cells, on which plate(s) would they most
likely be located? Explain your predictions.
- They’d be on the +pGLO plate, because the cells would survive long enough to be
transformed, because the transformation gives them the ability to survive the ampicillin.
3. Which plates should be compared to determine if any genetic transformation has occurred?
Why?
- I think the +pGLO plates should be compared against each other (one with arabinose,
one without). I say this because then it is a fair comparison, because they both have the
pGLO DNA.
4. What is meant by a control plate? What purpose does a control serve?
- A control plate exists to show a further difference in the final product of the experiment, a
meaning of comparison between different variables added. In this case, the difference
between arabinose and no arabinose, or ampicillin versus no ampicillin.
PAGE 43
1.
2. How much bacterial growth do you see on each plate, relatively speaking?
- On the +pGLO plate without arabinose, there’s some bacterial growth, but less dense
growth compared to the -pGLO plate with no ampcillin (presumably just e coli and the
broth?). On the +pGLO plate with arabinose, there’s more glowing bacterial growth it

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