1 / 16

Bacteriophage

Bacteriophage. Maddy and Megan . Introduction. Our goal was to isolate, purify and characterize our phage from a specific environmental sample. We are isolating the phage to be able to study the different genetic sequences. It may be useful in the study of bacterial diseases

pascha
Télécharger la présentation

Bacteriophage

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Bacteriophage Maddy and Megan

  2. Introduction • Our goal was to isolate, purify and characterize our phage from a specific environmental sample. • We are isolating the phage to be able to study the different genetic sequences. • It may be useful in the study of bacterial diseases • May also be useful with phage therapy

  3. Methods • We obtained a soil sample for phage isolation • We then enriched the soil sample • Then we harvested and prepared the sample in order to obtain plaques • Next we processed plaques • Then determined phage titer and made streak plates • Then purified the phage by titrating the sample two more times • We then titrated our sample a fourth time in order to get web plates and determined phage titer • Then we harvested a phage lysate by flooding our web plates and then usuing that we did more titration and made more plates • Then determined phage titer once again • Then we made more web plates using our MTL

  4. methods • We then flooding our web plates and filtered out liquid to make our HTL • Next we made plates to determine our phage titer • Then we used HTL to degrade bacterial DNA • Then we determined phage titer • Finally we isolated our phage genomic DNA • We then quantified our DNA using a spectrophotometer • Then we electrophoresed our DNA to take pictures • Then digested our phage genomic DNA with various enzymes and electrophoresed the samples to determine how the enzymes cut our DNA

  5. Godzilla • I found my original phage about a foot away from the river near the amphitheater. • It was at 44.8594555 latitude, -92.6218815 longitude • in a moist, 65mm depth, at 77 degrees

  6. Godzilla • I wasn’t able harvest any plaques from my soil sample so I then picked a plaque from one the Hudson phages • I originally found two different types, one was a small plaque and the other was a large one • During the process of purifying our phage, they both grew to be the same size • I then gave my other samples to Megan, because hers died

  7. Godzilla • My first titer was 1.14×10^6 pfu/mL • My second titer was 1.09×10^7 pfu/mL • My third titer was 6.2×10^7 pfu/mL • My fourth titer was 4.96×10^9 pfu/mL • I my final titer it was very low .061 pfu/mL in each one of my titer calculations

  8. Godzilla • My first phage genomic DNA electrophoresed sample was only cut by Hind3 • My second genomic DNA electrophoresed was cut by Nco1

  9. Godzilla • I used the website to look at the phage Salgado, and found mine was not cut the same • No enzymes related to my phage

  10. My phage measures about 25nm And the tail measures about 50nm

  11. Toulouse • My soil sample did not yield plaques • Used Hudson phages and did a titration • First titer: 9.87×104pfu/mL • Second titer: 1.494×107pfu/mL • Third titer: 2.7×104pfu/mL • Lost my phage during harvesting the phage lysate and titration • Adopted Maddy’s smaller phage

  12. Pictures of spot test and my 10-4 and 10-5 plate My titer was 1.265×109

  13. Then electrophoresed our DNA • Quantified DNA using spectrophotometer • .2545 μg/μL

  14. Digested our phage DNA and electrophoresed it My phage’s DNA was cut by Hind III and NcoI

  15. Conclusion • We think that our phage might be the same. • We were not able to find any other plaques that were similar to ours however ours were similar

More Related