IGEM:Virginia 2012/Protocols/Phage Replication Rate Assay

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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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# 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 freezer.
+
# 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 freezer after 3 hours and 6 hours.
+
# 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.

Revision as of 16:50, 29 May 2012

Contents

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 4 degree fridge.
  4. Put the bacteria/phage mixture in the incubator.
  5. Take 500 μl samples and put them in the 4 degree fridge 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!


References

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.

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