So on Monday, I had phage on all 5 plates from 10^0 to 10^-4. 10^0 and 10^-1 were webbed, and I had many on 10^-2. For the next part of my experiment I picked a plaque from 10^-4 and did a dilution once again. Tomorrow I will do the same thing, so that I can purify the phage. Tomorrow will be my 2nd purification, and next week on Monday will be my 3rd purification. I am so excited to be finally on my way to purifying and isolating a phage!
I am sorry that you did not find a phage, but I am glad you could adopt one. I am sure you will grow close to your phage as you will have many experiences with it for months to come.
Wednesday was proof that miracles do happen. On my last plate I found a phage! Now with my luck, it had contamination, but I was able to pick some phage from my plate. I diluted it to 10^-4 and will be ready to start the next procedure on Monday. I also use LB broth for my Msmeg. It is going to be interesting how that pans out, but it might work. We did not have any 7H9 top agar, so Dr. Adair said I should use the LB broth.
One thing that is confusing me is where the contamination is coming from. It is not the phage buffer because my control square was fine. The only other logical explanation is the top agar, but I am not sure if it is from the top agar, or the environment. I am going to just have to play around and work with it some more.
Hi Sea Phage Researchers,
Today after class I went and looked at my plaques, and I believe I have a phage! There was some contamination, but I think I can harvest some phage from my plaque. I obtained the phage from a flowerbed outside the BSB. I will start my purification on Monday after I have let my phages grow some. None of my other 6 samples showed promise of a phage. I was hoping to find some more phage in my other samples, but hopefully I will be able to harvest the phage I harvested from the BSB flowerbed. Next week should be promising.
On Wednesday, I prepared some Msmeg culture in 1X top agar to find some phage. I used my original 3 samples that I worked with at the beginning of the semester. I spotted my plates to 10^-3 to see what I can get. Hopefully I will begin purification of the phage today. I am especially hopeful about the sample that I obtained that was close to Waco. This was good loose soil for the phage find their host. I guess we will have to see today!
Another week of phage hunting starts tomorrow. I have 8 samples going right now. I have kept 4 samples going and 4 new samples I started on Wednesday. I have tried many different areas around Baylor such as a flower bed by Sid Rich, and a drain by the BSB. These seem promising since Arthrobacter thrive in harsh environments. I am hoping that the industrial chemicals in the flowerbed and the drain may attract Arthrobacter and as a result phage. C’mon phages!
I put 4 new samples in a spot test on Monday. I obtained my samples from my around the BSB and the SLC. I dug the dirt them from a field, a flower pot, and two flower beds. I diluted my samples only to 10^-3 and put them in 1X agar. I feel confident about a sample I took from the field in front of the SLC because there was standing water there for a while. Hopefully we get some phages tomorrow!
Sea Phage Researchers,
My sample yesterday still yielded no phage. I had some contamination on a plaque, but not where I put my soil samples. Therefore, I put my entire sample as negative for the Arthrobacter plaque. I filtered my third sample again and spotted a plaque to try this sample again. If this sample yields no phage, I will try an entirely new sample from around campus. Maybe this will yield better results.
Hi Sea Phage Researchers,
Since I finally learned how to use this blog today, I can actually post something about phages. Today Jeremy and I did not mix up our Arthrobacter bacteria samples. So that is positive. Because our recent trials have ended in either contamination or no activity at all, we used a different agar and a different host, and we also used the same agar and bacteria we used before to test our last sample of dirt that we acquired before lab started. I used a soil from my hometown in North Dallas and I also used a sample that I acquired just 30 minutes north from Waco. Hopefully these will capture some phage in the soil. I put all 3 samples in both of my agar plates just to grab some more info and retest one of my samples. I am excited to see what will happen on Monday!
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