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“ TymAbreu ”

“ TymAbreu ”. Discovered by Jessica Dahlke and Emma Sweet UW-River Falls. Background of Viruses.

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“ TymAbreu ”

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  1. “TymAbreu” Discovered by Jessica Dahlke and Emma Sweet UW-River Falls

  2. Background of Viruses For over a 100 years viruses have been in a scientific gray area; they are not quite living or dead materials, nor are they chemicals or poisons. Virus literally translates to “poison,” but it has been determined throughout the years that viruses are in fact not poisons. The association with poison came from viruses seeming to be connected to disease. Discoveries now have shown that though viruses do not possess all the requirements that it takes to be considered a living organism, they are biological beings that have amazing ability of utilizing true living organisms in aiding their replication and survival (Villarreal). Reference: Villarreal, Luis. "Are Viruses Alive?" Scientific American. (2004): 101. Web. 3 Dec. 2013.

  3. Why the name “TymAbreu”? • Emma’s original phage name was “Tyma” • Jessica’s original phage name was “Abreu” • After discovering they were the same phage, we just combined the two names to create “TymAbreu”

  4. Sample Source • Location: Hudson, WI • Karen Klyczek’s tomato garden soil • Amended with compost • GPS Coordinates: 44⁰ 59’ 4” N 92⁰ 45’ 16”W • When: September 21, 2013 at 9:30 am • Soil Condition: dry and loose

  5. Plaque Morphology • Size: • 1mm-2mm • Appearance: • Clear • No halo/ring • Titer of HTL: • Jessica’s sample: • 4.4 x 109pfu/mL • Emma’s sample: • 9.3 x 108pfu/mL FIGURE 1. JD HTL Prep 10-6 10-15-13 FIGURE 2. ES HTL Titer 10-5 10-22-13

  6. DNA Isolation Jessica’s Sample Concentration: 0.1945 µg/µL Yield: 19.45 µg (Now 14.45µg) A260/A280: 0.389/.213 = 1.826 Emma’s Sample Concentration: 0.08 µg/µL Yield: 8 µg A260/A280: .16/.09 =1.778

  7. Restriction Enzyme Results FIGURE 3. Jessica’s results from the first DNA digest. HindIII HaeIII EcoRI ClaI BamI Undigested DNA KB Ladder HindIII HaeIII EcoRI ClaI BamI Undigested DNA KB Ladder FIGURE 4. Emma’s results from the first DNA digest.

  8. Restriction Enzyme Results JD PstI JD SalI JD EcoRV JD NcoI 1KB Ladder ES NcoI ES EcoRV ES SalI ESPstI FIGURE 5. This is the second DNA digest results for both of the samples. They were set up to mirror image each other across the 1KB Ladder. This helped us determine that we had the same phage.

  9. Electron Microscope Results FIGURE 6. Emma’s EM image at 100K Head diameter: 50 nm Tail length: 106.25nm-118.75nm Type of phage: Siphoviridae FIGURE 7. Jessica’s EM image at 100K Head diameter: 50 nm Tail length: 81.25-103.13 Type of phage: Siphoviridae The size of the heads of the phage in both images are the same size, indicating that they are indeed the same phage. The range of tail length could have resulted from breakage while preparing the phage on the copper grid to be looked at under the electron microscope.

  10. Analysis Is TymAbreu unique? TymAbreu is unique when comparing the electron microscope images with other phages in the class. The head sizes of the other phages differ from TymAbreu’s and all the tail lengths differ, which is most likely due to breakage when preparing the phage on the copper grid to be observed using the electron microscope.

  11. Analysis (cont.) When comparing the electrophoresis gels, one can say that TymAbreu is not unique due to similar results throughout the class for both rounds of electrophoresis. PstI SalI EcoRV NcoI KB Ladder PstI SalI EcoRV NcoI HindIII HaeIII EcoRI ClaI BamI Undigested DNA KB Ladder FIGURE 8. Ashley Bue’s first round of electrophoresis. Compare to Figures 3 and 4. FIGURE 9. SJ and Sarah’s second round of electrophoresis. Compare to Figure 5. Figure 8 is just one example of the similarities found throughout the class for the results of the first electrophoresis. Figure 9 is a comparison of similar results for the second electrophoresis.

  12. The Journey Overall, we both thought this experience was rewarding. The process started out to be interesting and just an entirely new adventure. Towards the middle, it got frustrating at times because of errors, contamination, and other problems that occurred. We were also a little disappointed that we ended up having the same phage. But, once we saw our “phage babies” under the electron microscope, we were satisfied with the rewarding experience that was offered to us.

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