Download pGLO Lab Write up – Jauss Biology 3 - Parkway C-2

pGLO Lab Write up Part 1– Jauss Honors Biology 3
Do in lab notebook!
Title: pGlo Lab
Purpose: To transform E. coli bacteria by adding plasmids that allow the bacteria to glow green under UV
light in the presence of arabinose sugar and grow in the presence of the antibiotic, ampicillin.
Background:
1. What type of organism is being transformed?
2. Why is this organism a good choice for transformation?
3. Draw the pGLO plasmid and describe the 3 genes operating on the plasmid.
4. What was the origin of the GFP gene? How was it placed into the bacterial plasmid?
5. Describe the operation of the ara c gene.
6. Describe the operation of the GFP gene.
7. Are the BAD genes in the genetically engineered plasmid? Why or why not?
8. If the ara c gene is not switched on, will the bla gene still function? Why or why not?
9. What 3 substances are mixed in the agar solution. What is the purpose of each?
10. What is the purpose of the transformation solution?
11. What is the purpose of the ice followed by heat shock?
12. How will you determine if the bacteria have been transformed?
Materials: List everything you used in the lab today.
Procedure: List steps according to handout.
Day One Data: Put in regular notebook for now – as a draft
Observation of E. coli Starter Plate:
1. Draw Plate:
2. Number of colonies
3. Size Range of Colonies (in mm)
__________ to _____________
4. Color of Colonies?
5. Color of Colonies under UV light? (same color as 4)
6. Distribution of colonies on plate.
Observation of Plasmid:
7. Appearance of Plasmid
8. Color of plasmid under Normal light?
9. Color of plasmid under UV light? (same color as 8)