Lab 4: PRELAB

PRELAB ASSIGNMENT

• What will you be doing in lab and why?
• What should you end up with at the end of the lab period? Where will it be located?
• Prepare the calculations for your assembly reaction before coming to lab. First, complete the pre-lab quiz calculation, as described below. Next, calculate the number of picomoles present in each of your DNA samples and decide how you will set up your reactions (instructions and a tutorial video follow on the next pages).

PRELAB QUIZ

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.

You will need the expected sizes of the PCR products, as you determined for the prelab 3 quiz.

You can take the quiz in Canvas here (link opens in a new tab): Prelab 4 Quiz