Download p-glo lab 09-10

A Glowing Transformation – Honors Biology Lab (08-09)
Prelab
1. Using the Pearson Success.net website, read the introduction to “A
Glowing Transformation”, Lab 13 in Chapter 13.
2. Summarize your answers to the three prelab questions. BE SURE you
understand these answers.
3. What process was used to make the p-glo plasmid? Describe and/or
diagram it. REFERENCE PAGE IN BOOK ______________.
4. Predictions:
A. Plate on which the cells will be most like the original, nontransformed E. coli colonies?
LB________ LB/amp_______
LB amp/arabinose________
Explain your prediction:
B.
If there are any genetically transformed bacterial cells, o which
plate(s) would they most likely be located.
LB________ LB/amp_______
LB amp/arabinose
________
Explain your prediction:
C. Which plates should be compared to see if any genetic
transformation has occurred? Explain
Data Tables:
Make a data table in your notebook, using a ruler for each of the four dishes
Columns in the table:
1. Plate (LB, LB/amph, LB/amph/ara)
2. Relative number of colonies
3. Glow without UV light
4. Glow with UV light
5. Other observations
Analysis:
1. What did you see when you observed E. coli bacteria under UV
light on the original, non-transformed bacterial plate? Was it what you expected? Explain.
yet contain the gene for GFP.
2. What did you see when you observed the pGLO plasmid DNA under
UV light? Was it what you expected? Explain.
3. Did bacteria that were not mixed with the pGLO plasmid DNA grow
on the agar plate containing amphicillin? Explain.
4. Did bacteria that were mixed with pGLO plasmid DNA grow on
the plates containing amphicillin? Explain.
5. Do the colonies on both the plasmid plates glow under UVlight? (your answer should be no!!)
Why or why not? (this is the most important question of the lab! You must read about
and explain the role of the Arabinose operon to answer this question)
5. What are the key differences between arabinose operon different from the lactose operon
system in the textbook?
6. Describe exactly what is causing the bacterial colonies to glow.