I didn't deal with phage methods before. for that, I don't have the experience or the imagination so when I read "Isolate the top agar with the phage titre, then centrifuge" within procedures. I don't get it
This sentence is a little confusing. Can you describe what you are trying to do, or attach your protocol. The context might help explain what the sentence means.
The context is as follows:
Cells were plated on TSB top agar, then after overnight incubation, Top agar from plates with high phage titer was isolated and finally centrifuged.
I am assuming that what you are trying to do is to create a high titer phage stock using plate overlay?
So the routine procedure is to add around 5 ml of buffer or TSB liquid media to the top of the plate, let it sit for a 10-15 minutes to elute the phage and then either you can just remove the liquid and it should have high titer phage stock OR you can scrape the top agar off the plate using a spreader bar or something similar and put the contents of the top agar and liquid into a centrifuge tube and remove the top agar debris by centrifugation. The first is easier but the second sometimes gives you a slightly higher titer (but not always).
In both cases you should then sterilize the phage stock either by filter sterilization or addition of chloroform, or heat treatment depending upon the tolerance of your specific phage to those treatments.
I have another question:
I'm searching to order bacteriophage ϕ11 but I couldn't find obvious companies and the papers didn't mention the source. so I guess I miss something..
There are thousands and thousands (or maybe millions) of different phages. Companies only sell a very small subset of them, the ones that many people request and they can therefore make money from selling it. The other ones are generally either only found in a culture collection or from some lab working on that specific phage.
Assuming you are talking about Staph aureus ϕ11, you probably need to get from another Staph lab
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line