IGEM:Virginia 2012/Protocols/Phage Replication Rate Assay

Overview

Below is the procedure for measuring the rate of replication of a phage inside a certain organism. The bacteriophage and the organism are cultured together, and sample are taken and titered at 0, 3, and 6 hours.

Materials

• Liquid LB medium
• Bacterial culture
• Phage suspension

Procedure

1. Take 1 mL from the bacterial culture and add it to ~20 mL of liquid LB medium.
2. Add phage suspension to the bacterial culture.
3. Take a 500 μl sample and put it in the freezer.
4. Put the bacteria/phage mixture in the incubator.
5. Take 500 μl samples and put them in the freezer after 3 hours and 6 hours.
6. For each of the three samples, isolate the phage.
7. For each of the three samples, perform a plaque assay to determine the phage titer.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

Contact

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