Production of the Transient Phage Stock

Procedure 

  1. Cut 5–10 plaques with a sterile Pasteur pipette and place them in 1.5 ml sterile Eppendorf micro-centrifuge tubes.
  2. Add into 300 µl of Lambda buffer and incubate for 30 min at room temperature with intermittent gentle shaking every 10 min.
  3. Add 1:10 chloroform: lysate ratio with gentle shaking for 10 min at room temperature in order to elute the phages from the agar to lyse the bacterial cells.
  4. Incubate the mixture for 3 min in crushed ice.
  5. Cell debris is removed by centrifugation at 5000xg for 15 min at room temperature and the supernatant is transferred to a 1.5 ml sterile Eppendorf micro-centrifuge tube.

This method was originally published by Aldoori et al. 2015 with slight modification

Reference article

Barbosa, L. N., de Almada, A. F. B., Junior, J. A. S., Del Vechio, M. A., Bezerra, K., Espolador, G. F., … & Gonçalves, D. D. (2020). Bacteriophages’ action against mastitis-causing bactéria. Research, Society and Development, 9(10), e1849108541-e1849108541.

About the author

Hello there!
I'm Raphael Hans Lwesya, My true passion lies in the world of phage research and science communication. As a diligent phage researcher and an enthusiastic science communicator, I've founded "www.thephage.xyz," a platform dedicated to unraveling the fascinating universe of bacteriophages – viruses that specifically target microbes. My ultimate mission is to bridge the communication gap between the general public and the often intricate world of scientific concepts. I take pride in simplifying complex ideas, breaking them down into easily understandable pieces, and making cutting-edge phage-related research accessible to a wide audience. Thank you for visiting The Phage blog. If you have got any question or suggestion please drop it as a comment or via [email protected]

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