PhageIsabelle S, Ashley J, Brittany H

Intro• Phage are viruses that infects bacteria

• Phage can be used to treat diseases that cannot be treated by antibiotics

• Objective: to discover a unique phage to send in for sequencing. The phage they collect with different genomic sequences are put into an online database to compare to each other.

• Hypothesis: If we use standard biological techniques in the laboratory, then we will discover a unique phage to send in for sequencing.

Methods• Collect soil

• Enrich soil

• Harvest plaque

• Purify phage

• Harvest plate lysate (MTL & HTL)

• Isolate and purify phage DNA

• Quantify DNA

• Digest DNA

• Analyze DNA using electron microscopy

Musack (Ashley’s Phage)

• The name of my phage is Musack. The reason that I picked this name is because I am engaged and Musack will be my married last name. Musack is not a very common name but still not too complicated of a name either. Plus I thought it was a cool way to honor my new life with my soon to be husband and his family.

• The soil sample that I collected did not produce any plagues, even after re-enrichment. This phage came from a plaque that I picked from the “Hudson phage” plate that was labeled 10^-2. My initials are on the plaque that I picked on that plate.

• Plaque size: Relatively small

• My HTL titer was 4.285×10^9 pfu/mL

• My MTL titer was 3.285×10^9 pfu/mL

Musack

• Yield of DNA extraction was .1095 µg/µL within a 100 µL sample

• The absorption rating in 260 light was .219

• The absorption rating in 280 it was .136 for a ratio of 1.610

• My phage, when cut with Nco I, looks similar to the phage Bernie and the phage Salgoto. They both have the double band in about the right place and in Salgoto there is a multiple banded section towards the end that looks similar to the end of the banded region in my gel.

Measurements of Phage

• Diameter of head : 40 nm

• Length of tail : 110 nm

Sadie (Isabelle’s Phage)

• I named my phage Sadie because I am obsessed with my dog, and her name is Sadie.

• I collected soil in River Falls but it did not produce any plaques. I then picked from the 10^-2 Hudson Phage plate.

• Titer of MTL : 1.42 x 10^9 pfu/mL

• Titer of HTL : 7.9 x 10^9 pfu/mL

Sadie

• Yield of DNA: .123 µg/µL

• The absorption in 260: .246

• The absorption in 280: .130

• .246/.130 = 1.89

Measurement of Phage

• Diameter of head : 25 nm

• Length of tail : 105 nm

Hans4232 (Brittany’s Phage)

• I named my phage Hans4232 because it was my school ID from Morris.

• Since the soil I collected didn’t produce any plaques, I picked from the 10^-2 Hudson Phage plate.

Measurement of Phage• Diameter of head : 45 nm

• Length of tail : 95nm

Conclusion

• Ashley : No

• Isabelle : No

• Brittany : No