Studier Lysate Prep

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Summary

How to make a lysate from a plaque preparation.

Protocol

  1. Add 5mL fresh overnight (BL21 if wild-type T7) into 15 mL of T broth (125 mL flask)
  2. Add a single plaque to above
  3. Shake at 30°C until lysis is visually apparent
  4. As soon as lysis is observed and appears to be complete, add 1g of NaCL to the flask and dissolve by shaking.
    • Studier has observed that leaving released phage in the cell debris causes a reduction in the kinetics of phage binding (personal communication). It becomes difficult to get good time courses in this case. It is recommended when making a large stock that will be used in timing experiments, that you check that most of your phage adsorb to the cell in the first 30 seconds.

References

Studier, FW Virology 39:562 (1969) Link: Pubmed

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