Lab 4: LAB ASSIGNMENTS

PRELAB QUIZ

Visit Canvas to take the quiz. The quiz questions are also listed below for your convenience:

Question 1

What will you be simulating in lab? What is the purpose of those experiments? (Provide 2-3 sentences).

Question 2

In order to set up a Gibson assembly reaction, you will have to calculate DNA molarities. As a first step, calculate the molecular weights of vector (pETblue2) and insert (HCAII) in g/mol. To determine the MW of your DNA, assume that one DNA basepair = 650 g/mol. This is an estimate, but it is okay to use this number to calculate the molar amounts of your DNA.

Note: to solve this problem you will need the sizes of the PCR products, as you determined in Lab 2.

Question 3

This lab includes a safety hazard. Describe the hazard and steps you would take in a lab to protect yourself and your colleagues.

IN-LAB ACTIVITIES

  • This week we will have the second of two TA Talks. One of your TAs will introduce themselves more thoroughly by telling you a bit about how they got interested in science, how their graduate school career is progressing, and what they are currently researching. The talk will last ~15 minutes and then you will have an opportunity to ask questions about their research, graduate school, etc.
  • You will then be randomly assigned to a breakout group with a lab partner, and you will be provided the link to a Google doc. You and your partner will work together to complete it. It will include:
    • A table where you will outline each step in the protocol, state its purpose, and write down any questions you have. Each pair is required to ask a TA at least one question about the protocol.
    • Completing the calculations for setting up the Gibson Assembly reactions.
    • Complete a virtual bacterial transformation lab.
    • Consider some approaches to troubleshooting common problems.
  • Before leaving BBCU, meet with a TA to ask them at least one question about the lab. They will also check your GA reaction calculations.
  • Complete the lab/partner/self evaluation Google form (link will be provided).
  • Get a sharable link to your Google doc and submit it to Canvas.

License

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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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