Team:BYU Provo/Notebook/LargePhage/Springexp/Period1/Exp/Team:BYU Provo/UV Selection Test


Large Phage May - June Notebook: Experiments


UV Selection Test

I) Purpose

The purpose of this experiment is to subject small samples of phage to varying doses of UV light. This experiment is thought to work by selecting against phage with only one copy of each gene. When critical genes are knocked out of these phage, the phage cannot replicate themselves. Phage with multiple genomes (and thus, larger capsids) are much more resistant to UV light because they have multiple copies of each gene.

II) Expected Outcome

We predict that phage more resistant to mutation and UV light will be the ones that will survive the irradiation the best.

III) Materials

Liquid high-titer phage lysate
Overnight culture of E. coli
UV light (we used the UV light inside a BSL-2 hood)

IV) Methods

1. Under the hood, divide a piece of paraflim into sections, labeling each section with a time for how long that sample will be subjected to UV light. (We usually do 1 minute increments)
2. Place 20 uL from the phage stock on each section of parafilm.
3. Start the UV light for the incremental time amount. When the time is up, turn the light off.
4. Remove 10 uL from the first section and place it in 0.5 mL of E. coli in a test tube.
5. Start the UV light again, and repeat steps 3-5.
6. Plate out all infected E. coli samples in LB top agar.

V) Results