IGEM:Virginia 2012/Protocols/Phage Replication Rate Assay: Difference between revisions
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(New page: ==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 ta...) |
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==Overview== | ==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 | 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 samples are taken and titered at 0, 3, and 6 hours. | ||
==Materials== | ==Materials== | ||
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# Take 1 mL from the bacterial culture and add it to ~20 mL of liquid LB medium. | # Take 1 mL from the bacterial culture and add it to ~20 mL of liquid LB medium. | ||
# Add phage suspension to the bacterial culture. | # Add phage suspension to the bacterial culture. | ||
# Take a 500 μl sample and put it in the | # Take a 500 μl sample and put it in the 4 degree fridge. | ||
# Put the bacteria/phage mixture in the incubator. | # Put the bacteria/phage mixture in the incubator. | ||
# Take 500 μl samples and put them in the | # Take 500 μl samples and put them in the 4 degree fridge after 3 hours and 6 hours. | ||
# For each of the three samples, [http://openwetware.org/wiki/IGEM:Virginia_2012/Protocols/Phage_Isolation isolate the phage]. | # For each of the three samples, [http://openwetware.org/wiki/IGEM:Virginia_2012/Protocols/Phage_Isolation isolate the phage]. | ||
# For each of the three samples, perform a [http://openwetware.org/wiki/IGEM:Virginia_2012/Protocols/Plaque_Assay plaque assay] to determine the phage titer. | # For each of the three samples, perform a [http://openwetware.org/wiki/IGEM:Virginia_2012/Protocols/Plaque_Assay plaque assay] to determine the phage titer. | ||
Latest revision as of 20:32, 29 May 2012
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 samples are taken and titered at 0, 3, and 6 hours.
Materials
- Liquid LB medium
- Bacterial culture
- Phage suspension
Procedure
- Take 1 mL from the bacterial culture and add it to ~20 mL of liquid LB medium.
- Add phage suspension to the bacterial culture.
- Take a 500 μl sample and put it in the 4 degree fridge.
- Put the bacteria/phage mixture in the incubator.
- Take 500 μl samples and put them in the 4 degree fridge after 3 hours and 6 hours.
- For each of the three samples, isolate the phage.
- 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!
References
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
