Lab 3: WRITE UP

There is no mini lab report this week; instead, you need to answer the following questions about your at-home experiments. You do not need to format your responses as you would a lab report; just answer the questions.

Questions from Activity 1: Sickle Cell Anemia Detection and Simulation

  • For the wild-type beta-globulin sequence shown in the protocol, how many DNA fragments are present after digestion with DdeI?
  • For the mutant beta-globulin sequence, how many DNA fragments are present after digestion with DdeI? If the number of DNA fragments is different than that of the wild-type, explain why.
  • Count the number of base pairs for each fragment, for wild type and mutant. (If a base is not paired because of a sticky end, do not count it.)
    • For the DdeI-digested, wild-type beta-globulin sequence, how many base pairs are in each fragment?
    • For the DdeI-digested, mutant beta-globulin sequence, how many base pairs are in each fragment?
  • On the basis of fragment size, how can the difference between the wild-type sequence and the homozygous mutant sequence be recognized?
  • What fragments would be present following DdeI digestion of a sample from someone with a heterozygous beta-globulin genotype?
  • Insert a photo of your completed gel diagram from protocol Figure 4.
  • Based on your gel diagram, what is the genotype of the unknown sample from the newborn? Explain your answer.

Questions from Activity 2: Mini Gel Electrophoresis of Dyes

  • Insert a photo of your completed gel
  • Which combination of dyes make up each of the two unknown samples?
  • What causes different banding patterns for different samples? What do the banding patterns indicate?
  • Consider this experiment compared to the simulated HCAII experiment we did last week. How was this at-home lab similar to and different from doing gel electrophoresis in a research lab setting? Include two similarities and two differences.
  • It is actually quite impressive that you were able to do gel electrophoresis at home for only $22 (batteries not included). Can you imagine any real-world applications for this sort of technology? Describe one idea in detail.

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Biochemistry 551 (Online Version) Lab Manual by Lynne PROST is licensed under a Creative Commons Attribution 4.0 International License, except where otherwise noted.

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