Enrichment Cultures for Phage Isolation

Enrichment Cultures for phage isolation

1. Add 0.5 ml of a saturated M. smeg. culture to 5ml enrichment media in a large culture tube.

2. Add < 0.5 gm of environmental sample (typically dirt) to culture tube.

3. Let grow for 1-2 days with shaking at 37°C.

4. Remove tubes from incubator and let sit for a few minutes, allowing particulate material to settle out.

5. Sterile filter 0.5-1.0 ml of culture into a sterile, labeled eppendorf tube.

6. Plate filtrate to a lawn of M. smeg. to determine if phage are present.

Notes:

1. dispose of remaining culture to biohazard waste.

2. After plating filtrate, store the remaining undiluted filtrate at 4°C—you will need this to purify your phage.