IBEHS 2P03 Wet Lab 2
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Apr 3, 2024
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IBEHS 2P03 Wet Lab 2
Owaiz Merchant
400199784
Due: February 4
th
2023
1. Are the colonies on your plates countable? Why or why not? If your plates are not countable,
what could you try if you were to repeat the experiment for a second time? (2-4 sentences).
The colonies on my plate are countable. I am able to count it by looking at the holes made in the agar gel. However, there is not many colonies on the plate. In order to change this next time, I could add a higher concentration of the bacteria.
2. For the next series of questions, assume every colony on your dilution plates is descended
from one colony forming unit (CFU
a.
Which dilution tube did you plate in your series?
I used the 3
rd
tube in a 100x dilution series.
b.
How many CFU did you seed onto your dilution plates?
I seeded 39.7 CFU on the first plate and 27.79 CFU on the second plate.
c. What is the concentration of your selected dilution tube? Can this be calculated from
your plates?
The concentration of the dilution tube is 79.4 CFU/mL. This can be calculated by looking at the plates by seeing how many small holes are on the plate as each represents a colony forming unit.
d. What is the concentration of the original, stock solution of cells you were provided?
Can this number be calculated from your plates (1-2 sentences)?
The concentration from the original stock solution of cells is 7.94*10^7 CFU/mL. This can not be calculated as it all depends on how much volume is placed on the gel and how diluted the solution was made.
3. In the last lab, you used an equation provided by your TAs to estimate the concentration of the
original stock solution, and then used this number to plan your dilution series to plate
countable colonies.
Does the number in 2d match the estimated concentration you obtained using the plate reader
number? Provide a potential explanation for why the numbers match or why they are different
(1-2 sentences).
The numbers do match as the amount of holes counted on the plate equal to the CFU calculated above.
4. After your gel electrophoresis is complete. Your TA will take an image of the gel and upload it
to Avenue. You need to annotate the gel image, labelling the lanes with the lane number and
identifying what is in the lane. You need to identify the ladder as well. You need to number your
image (e.g. Figure 1) and include a descriptive caption below your image. Lastly, answer the
following questions regarding your gel:
a. Was your PCR successful? How do you know? If not, list at least 3 reason what might
have gone wrong.
Figure 1: Control on the left, miniprep in the middle
b. Was your miniprep successful? How do you know? If not, list at least 3 reasons what
might have gone wrong.
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