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

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