Studier Lysate Prep
How to make a lysate from a plaque preparation.
- Add 5mL fresh overnight (BL21 if wild-type T7) into 15 mL of T broth (125 mL flask)
- Add a single plaque to above
- Shake at 30°C until lysis is visually apparent
- 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.
Studier, FW Virology 39:562 (1969) Link: Pubmed