Donald P. Breakwell:Phage Enrichment

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This protocol has been copied and adapted from the HHMI Science Education Alliance protocol.

A. Prepare the culture.

 1.  Grow a turbid culture of the host bacterium in an appropriate medium.  Ideally, the culture should be late log to early stationary phase.
 2.  In a sterile 250mL erlenmeyer flask, add 25mL fresh growth medium supplemented with 1M CaCl2 to a concentration of at least 50 mM. Inoculate it with 5 mL of the turbid culture.

B. Collect a soil sample from an appropriate site.

 1.  Use a 15-ml conical tube.  Label it with the soil identification number, your name, and the date.
 2.  Record other pertinent sampling details on a Phage Collection sheet.

C. Inoculate the enrichment

 1.  To the 250mL erlenmeyer flask containing inoculated growth medium, add 0.5 g soil (about 0.5 teaspoons).  This need not be too accurate.

D. Incubate in a shaking incubator (200+ rpm) at an appropriate growth temperature.