Hi Ben! Great job on the presentation! Are there specific strains or types of bacteria would the phages be able to interact with and potentially kill? Or is it a broad spectrum?
Thanks! We already know through our plaque assay that our phage infects M. smeg bacteria but more tests are needed to determine whether or not our phage has a broad spectrum of hosts.
Sure! So we exposed the isolated DNA of our phage individually to each of these restriction enzymes: HindIII, EcoRI, BamHI, CalI, HaeIII, and SalI. Then we ran all six test groups on a 8% agarose gel at 120v for ~30 minutes. By observing the unique cuts made on the gel by the different enzymes, we were able to put the data from each enzyme cut into a phage database. The database then showed us what phage were most similar to the data we put into the website, and 19/20 of the phage that were most similar belonged to the A1 subcluster.
We first collected our soil sample, and then used a process called enrichment which creates what is known as phage lysate. By mixing our soil sample with phage nutrients (bacteria) we hoped to create a high concentration of phage in the solution so that when they are filtered and added to a lawn of the bacteria on an agar plate, they will successfully create plaques. These plaques are were the phage devour the bacteria, and we can use them to isolate and conduct serial dilutions on the resulting lysate.
For sure! Anywhere there is bacteria there will be phage. A side purpose of this lab is to show that even dirt has endless amounts of phage. With that said, I can think of much more dirty/bacteria-ridden places that would have more phage to find.
Hi Ben! Great job on the presentation! Are there specific strains or types of bacteria would the phages be able to interact with and potentially kill? Or is it a broad spectrum?
Thanks! We already know through our plaque assay that our phage infects M. smeg bacteria but more tests are needed to determine whether or not our phage has a broad spectrum of hosts.
Can you explain in more detail how the restriction enzyme digestion products are analyzed to determine the phage type?
Sure! So we exposed the isolated DNA of our phage individually to each of these restriction enzymes: HindIII, EcoRI, BamHI, CalI, HaeIII, and SalI. Then we ran all six test groups on a 8% agarose gel at 120v for ~30 minutes. By observing the unique cuts made on the gel by the different enzymes, we were able to put the data from each enzyme cut into a phage database. The database then showed us what phage were most similar to the data we put into the website, and 19/20 of the phage that were most similar belonged to the A1 subcluster.
How do you isolate the bacteria of the phage?
We first collected our soil sample, and then used a process called enrichment which creates what is known as phage lysate. By mixing our soil sample with phage nutrients (bacteria) we hoped to create a high concentration of phage in the solution so that when they are filtered and added to a lawn of the bacteria on an agar plate, they will successfully create plaques. These plaques are were the phage devour the bacteria, and we can use them to isolate and conduct serial dilutions on the resulting lysate.
Are there other and better methods of finding phages?
For sure! Anywhere there is bacteria there will be phage. A side purpose of this lab is to show that even dirt has endless amounts of phage. With that said, I can think of much more dirty/bacteria-ridden places that would have more phage to find.
what were the limitations to this study?