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.