Rao:Transduction

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Overview

Generalized transduction is another powerful tool in bacterial genetics. In strain construction, generalized transducing bacteriophages are an indispensable tool. Below is an adaptation of the great protocol in the Sauer lab protocol.

The generalized transducing phages used in the Rao lab are P1vir (E. coli), P22HT (S. typhimurium), and PBS1 (B. subtilis). These are derivatives of temperate bacteriophages which have been mutated to only allow for lytic growth.

Materials

  • Glass Pasteur pipets. DO NOT USE YOUR PIPETMAN!!! (You will need to calibrate your pipetting - ie How many drops is 100 uL?)
  • Phage lysate
  • Culture tubes (use the plastic ones... less chance of phage contamination)

Method

P1vir

Use the Sauer Lab protocol: Sauer:P1vir_phage_transduction

P22HT

More to come

PBS1

More to come

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