Donald P. Breakwell:Phage Enrichment
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.