Lab 3: 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?
- You will encounter a chemical hazard this week in lab. Describe the chemical, the potential risks, and steps you will take to protect yourself and your colleagues.
- In this lab, you will be preparing 1% agarose gels by adding solid agarose to a buffer and then melting it. If you are going to prepare 50 mL of agarose solution, how much agarose (in grams) will you need?
PRELAB QUIZ
This week you will be running your HCAII and pETblue2 PCR products from last week on an agarose gel. About what size do you expect the successful products to be?
1) HCAII:
a) 780 bp
b) 840 bp
c) 1,000 bp
2) pETblue2:
a) 3550 bp
b) 3650 bp
c) 3700 bp
To answer these questions, consider:
- the pETblue2 vector map, available here (pdf opens in a new tab): pETblue2 vector map
- the HCAII full sequence, which can be found in the lab manual appendix: Appendix 1: Information on HCAII
The map shows that pETblue2 is 3653 bp in size. However, you did not amplify the entire plasmid. You need to think about exactly where the primers annealed in order to determine exactly how many bases were amplified. In addition, you added insert sequence to your primers, which will add bases to your final product.
Similarly, when considering the HCAII product, remember that your primers added additional bases to the product.
You can take the quiz in Canvas here (link opens in a new tab): Prelab 3 Quiz
Here is a video explaining Prelab 3 Quiz: