Finding and Isolating Phages

[@noutinf.michodigni7943]

Hi Dr. Sabrina and thanks for making public this practical knowledge. I kindly request that your expound on this detailed protocol by: 1) highlighting the name of the buffer(s) used for both isolation and storage of phages in your lab; 2) describing the recipe of the buffer(s), especially their final concentrations and the final pH value(s) of the buffer(s). Regards!

[@BCMTAILORLabs]

Yes, this was modified at the link above. Thank you for your suggestions.

[@emiliankozma3772]

Thank you for very nice video demonstration. I would appreciate to receive information that noutinf.michodigni7943 has asked .

[@synestthesium412]

I used this method to isolate T4 Bacteriophage from cow slurry in Dumfries, Scotland!

[@charlyapain620]

Hey, great video. I was wondering if you had any assays/protocols that involve evolving phages and bacteria together?

[@forbesavila8827]

Love the video. But why do you need the hard agar? Why not just use the soft agar? Thanks in advanced good luck!

[@ohoudahmad8273]

thank you for sharing this useful video. but i have one question if you don,t mind, how were bacteria selected?

[@desbonafide]

Thank you for this video!! You made my protocol much easier

[@monikafelegyhazi529]

Hello ! I have bacterial biofilm infection. Staphylococcus biofilm. Do you have phage produts ? Can I send a sample for examination ? Antibiotics doesn't work. I would like a phage therapy treatment.

[@Fidelanne]

Hi what's your location?

[@monikafelegyhazi529]

@@Fidelanne Hungary, in Europa.

[@Fidelanne]

@@monikafelegyhazi529 Okay please.

[@Fidelanne]

You are a bit out of reach logistically

[@monikafelegyhazi529]

@@Fidelanne Do you have phages ? Do you use phage to treat people ?

[@TeamKitrinos2017]

when I discovered bacteriophage was a thing I started to be super fascinated in viruses

[@markwolvers6298]

Very inspiring video to see how easy this is, and how much Dr. Green enjoys her work... *wiggle wiggle* Now I want to go hunt for phages as well! There was one aspect that slightly confused me, which was the ending of the video: Why did you cut the last sentence short and suddenly show an MRSA plate incubated with pig fecal lysate? Are all the transluscent spots there plaques caused by phage-induced lysis too? Those assumed plaques seem very homogeneous, which raises another question: Do you normally isolate only a single strain of phage from your lysates? And are filtrates and residues ever shotgun sequenced to explore whether the protocols used induce any form of selection bias?

[@BCMTAILORLabs]

Sorry to confuse. Great comment! That last plate was the result of the plaque isolated and replated to make a plate lysate. That is why the plaques were homogeneous. They were all derived from a single plaque! The plate is then used for further propagation and purification of the phage. Usually we get a lot of different looking phage plaques. But because you use only one isolation strain you are going to be biased.

[@markwolvers6298]

@@BCMTAILORLabs aha! So your overnight culture of bacteria was MRSA, and at the end you show the endresult of all your work? I assumed it was E. coli since you mentioned that species in the intro, but now I understand that was just an anecdote. I like the sound of that, a lot of different looking plaques, so you are peeking into a microbial jungle! Exciting.

[@weitingLin66]

Really nice video!

[@TamannaWara]

Thank you so much for this video! You are a godsend. I have a presentation tomorrow on use of phage therapy against anthrax as potential means of bioterrorism threat. & I am starting right now 🤣 & your video just about cleared everything!

[@nixbondi]

Isn't anthrax a virus and phages are bacteria killing viruses not virus killing viruses, right?

[@TamannaWara]

Yes. So how this essentially works is, as we know Bacillus anthracis is a bacterium that causes anthrax disease and phages are viruses that kill bacteria. Therefore phage therapy is used to introduce bacteriophages in the host (human with anthrax) to attach & lyse with B.anthracis' cell wall, they penetrate and hijacks the entire biological machinery of the bacteria. The phage then enters either undergoes the lytic or lysogenic cycle. The lytic cycle involves the reproduction of more bacteriophages after successfully infecting the host cell to manufacture more viruses; the viruses then burst out of the cell & the parent cell disintegrates (the OG B.anthrax bacteria that was hijacked. So instead of more B.anthracis being reproduced from one cell, you have mini phages coming out. Bacteriophages are inturn harmeless to humans. In the lysogenic cycle, the phage incorporates its viral genome (DNA) into the host cell (anthrax's chromosome) genome, infecting it from within. This allows the phage DNA (prophage) to be copied and passed on along with the cell's own DNA. Hence making anthrax say goodbye to its ability to produce more copies of its-own-self. Also, apart from genetically engineering phages against B.anthracis, Phages active against B. anthracis (both lytic and lysogenic) are widespread in the environment and have been isolated from soil, carcasses, feces, sewage, and the intestinal tract of the earthworm.

[@AnFarm-fx5ls]

which media did you use to isolate phages?

[@Luis.mp4]

when will phages become a worldwide medicine and get out of testing phase?

[@stephentrueman4843]

Phage therapy is used in europe

[@manishuaibu280]

What is the name of that filter

[@sashikantaparida9763]

HI IT IS A VERY NICE VIDEO BUT THE DETAILED PROTOCOL LINK IS NOT OPENING

[@BCMTAILORLabs]

Will fix

[@BCMTAILORLabs]

The link is through google docs. It seems to be working. We can send you a protocol if necessary. If you are on twitter @bcmtailorlabs direct message us.

[@kimanipatrick]

I have 3 Bacteria in mind that i have not been successful with finding phages for. How can i contact your for ideas?...

[@aguerrero1603]

Hi! I have a question. So let's say I have my bacteria and probably phages in my sample, but it seems there are very few plaques... If I want to get more phages, does it affect when I isolate my bacteria from time to time for it not to die ? Or do I need a new sample?

[@arulross70]

Hey do you think you could replicate this process for another virus? Like one that infects humans?

[@BCMTAILORLabs]

This is different with human cells. Our expertise is only bacterial viruses.

[@arulross70]

@@BCMTAILORLabs Couldn't you just take an infected sample and find the abundant viruses that should be there? Just curious.

[@venupal]

Hi.mam.will u plz help me in developing phage against vibrio.plz mam

[@mrtoco4474]

Can you make a phage for Cutibacterium acnes please please please is it possible to find it ? 😫😭😭😭😭😭

[@diamondeyes7274]

how did you do it in easy way??? i have been done for many time but, always failed. this is really aspiring teaching video..

[@mekkihalima320]

It is the same thing for me

[@moulaymhammedakermi3410]

Hi....whate kind of baffer you used to isolat phage?????

[@moulaymhammedakermi3410]

Î' am hope you give me a complet and easy protocle to isolat this phage.....

[@marwak9946]

What kind of buffer do u use ?? How to prepare it plez .. i have a problem with the palques formation in order to calculat the phage concentration in the pahge filtrat .. i do have a phage but it doesn’t form an individual plaques in the plate although i ttried so many timmes to preform the test ..thanx a lot

[@BCMTAILORLabs]

The buffer is in the recipe. Detailed protocol link bit.ly/397acAi

[@marwak9946]

And I’ll be greatfull if u write down the ingredients of the buffer

[@BCMTAILORLabs]

The buffer is here bit.ly/397acAi

[@thekidsnextdoorsneighbor3645]

I love it when nerds nerd 🥹😂

[@kathrynion]

Indentations?

[@BCMTAILORLabs]

We were referring to the difference of the clearing of a bacterial lawn verses an artifact caused by an air bubble. The best way to know is to just pick anything that looks like a plaque and replate it to make another lysate.

[@aniszulaika9641]

Why is it hard to filter the sample in the beginning of the procedure?

[@ameslee2153]

I believe it is due to the stools and other large particles (even bacteria) blocking the filter pores as centrifuging doesn’t always “pull” all of them down.