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

Icon for the Creative Commons Attribution 4.0 International License

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

Share This Book