Reagan Shaw, Mark Nino Doria, Lillian Cole, Christopher Kinzel, Ryanna Kennedy

October 19, 2023

Post Lab Transformation

1.
Plate: Prediction: Reason: Observed Result:

LB -plasmid Yes The plate would only contain N/A

the bacteria, not the ampicillin,
which would prevent growth

LB +plasmid Yes The plate only contains the N/A

bacteria and plasmid, which
would also help the bacteria to
grow.

LB/AMP No The plate would contain the N/A

-plasmid ampicillin but without the
plasmid, so the ampicillin kills
the bacteria with nothing to
prevent it.

LB/AMP Yes The plate would contain both Yes

+plasmid the plasmid and ampicillin.
While ampicillin would kill the
bacteria, the plasmid would
make the bacteria be able to
resist, and lead to its growth.
2. Plates have been observed.
3. Our observed results for the LB/AMP +plasmid plate aligned with our prediction that the
bacteria would grow. The other plates were not tested so therefore there are no observed
results for them.
4. LB/AMP+plasmid Plate 1: 6 colonies
LB/AMP+plasmid Plate 2: 15 colonies
5.
6. We selected a plasmid that contains a gene for ampicillin resistance so that after
transformation the bacteria that took up the plasmid can be distinguished from those that
did not by plating the bacteria on a medium containing ampicillin.
7. The phenotype of the transformed colonies tells us that the plasmid DNA has been
successfully taken up by the E. coli and is being expressed in the transformed cells.
8. One would first observe the ampicillin (LB/AMP) plate to determine if the transformation
was successful. You would observe the ampicillin plate because the e coli would not have
survived on a plate without it.
9. a) The total mass of plasmid used in pVIB is 0.05 micrograms.
b) The total volume of cell suspension prepared is 510 microliters.
c) The fraction of the total cell suspension that was spread on the plate is 10/51
microliters.
d) The mass of plasmid in the cell suspension spread was 3.921 micrograms/microliters.
e) The number of colonies per microgram of plasmid DNA in plate 1 is
1.530221882172915 × 100 and 4.08059168579444 × 100 in plate 2.
10. The factors that might influence the transformation efficiency are the concentration of
ampicillin, temperature, and procedure. Ampicillin is used for intentionally choosing to
obliterate E. Coli which didn’t possess an ampicillin resistance gene. Depending on the
amount of ampicillin added, the growth of the bacteria varies. Additionally, temperature
affects the competence of the transformation by exposing E. Coli to inconsistent
conditions could negatively impact the result. Moreover, conducting a procedure with the
error or masterly overall determines the transformation efficiency.
11. Useful human protein can be produced inside another species because of recombinant
DNA technology which allows for human genes to be isolated and inserted into the
common bacterium of a different species. This allows for large amounts of the protein
encoded in the gene to be produced with the help of the bacterium. This is important
because it allows scientists to do research to help humans in a much more efficient and
effective way since bacteria is smaller than humans, and can divide and reproduce faster.
There is also no ethical concern when using bacteria for research which is helpful. One
example that shows the importance of this is insulin because insulin is manufactured in
large quantities within bacteria.