Lab 8: PRELAB
PRELAB ASSIGNMENT
- What will you be doing in lab and why?
- You will encounter a chemical hazard this week in lab. Describe the chemical, the potential risks, and steps you will take to protect yourself.
- You will be running two gels this week. Each gel will be cast with 10 wells for loading samples and you will need to run one lane of protein marker on each gel. Since you cannot run everything you collected, you need to determine which samples will give you the most information.
- Keep in mind that your goals are, generally, to determine whether or not HCAII was overexpressed by the E. coli, and whether or not your column successfully purified HCAII.
- Before coming to lab, draw a picture of the two gels you plan to run and label all of the lanes with the samples you will be loading into them.
HINT: Load the protein markers differently in your two gels so that you can distinguish between the two (i.e. put the ladder on one end for gel 1, and in the center for gel 2).
PRELAB QUIZ
- After collecting your purified protein from the dialysis tubing, you will measure the A280 in order to determine protein concentration. As we will learn in lecture, blanking is very important when using spectrophotometric techniques. What will you use as a blank for this measurement in lab (Hint: what happens during dialysis)?
- If you assemble your gel apparatus and find that it leaks, describe one way you can try to fix it.
- Which is likely to polymerize faster: Resolving gel (bottom, poured first), or Stacking gel (top, poured second)
Link to prelab quiz (will open in new tab): Prelab 8 Quiz