Copy of 3. Pre-lab quiz questions

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Biology

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Feb 20, 2024

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Bio 153: Spring 2023 Lab 3: Purification 1 or 2 During the first 10 minutes of lab 3, you will be asked 5 of the following questions (2.5 pts each). You may bring handwritten notes (on paper) to use during the pre-lab quiz. No other resources will be permitted. 1. What is the purpose of the purification phase of this research project? ensure that you end up with a phage sample that contains only one kind of phage 2. What features of your plaques should you record in your lab notebook? Record the size, shape, and appearance of your plaques 3. How will you know from your purification plates if your serial dilutions were performed correctly? If you see the most phage on your most concentrated plate 4. What is the purpose of making serial dilutions of your picked plaques? To calculate the concentration of phages in the solution. 5. After picking a plaque and making serial dilutions, what is the last step? perform the plaque assay for all samples 6. When purifying a phage, it is important to understand that phage particles diffuse within the agar plate. Therefore, phages from a nearby plaque could diffuse into your selected plaque. How can you minimize the presence of other unwanted phages during the purification process? you should pick from a well-isolated plaque taken from a highly dilute phage sample 7. According to classic microbiology techniques, how many rounds of purification should be performed? three rounds of purification should be performed to ensure a single clonal phage population 8. What can happen if too many rounds of purification are performed? they provide your phage with the opportunity to accumulate mutations when its genome is replicated within the bacterial host 9. How will you know when you have successfully purified your phage? all the phage particles in your sample should be genetic clones of each other 10. What is the name of the host bacteria we are using in this project? Microbacterium foliorum

Bio 153: Spring 2023 11. What is a plaque? Plaques are formed on bacterial lawns when one phage particle infects and destroys a bacterial cell, releasing more phage particles. 12. Under what circumstances are 1-2 rounds of purification sufficient? if there is only a single plaque on the plate 13. If you have a plate with two different sizes/shapes of plaques on it, how can you determine if you have different phages or one phage that happens to make different plaques? pick a plaque for each morphology, serially dilute each sample, and plate dilutions for each sample using a plaque assay to see if these morphologies persist or can be separated. If the morphologies separate , you’ve shown that there are two phages. 14. What are two tips you should follow to take a good plate photograph? Remove lid, face agar side up and Place plate on dark surface 15. What is the difference between the first and second stop with a micropipettor? The first stop of a micropipette is used to collect fluid in the tip whereas the second stop is used to completely discharge the fluid. 16. From the lab math packet: What is the typical temperature of a refrigerator? 4*C 17. What is the typical human body temperature? 37*C 18. Top agar is typically kept at around 60 ℃ . How does this compare to room temperature and human body temperature? Room temperature is about 20*C, which is 40*C lower than the top agar. Human body temperature is about 37*C, which is 27*C lower than the top agar. 19. How many times more concentrated is a 10 -2 dilution compared with a 10 -4 dilution? a 10^-2 dilution is 100 times more concentrated than a 10^-4 dilution 20. How many times more concentrated is a 10 -1 dilution compared with a 10 -4 dilution? a 10^-1 dilution is 1,000 times more concentrated than a 10^-4 dilution

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