2. Centrifuge 1–2 ml samples of the overnight culture. 3. Resuspend the pellets in 350 μl of STET buffer (10mM Tris-HCl, pH 8.0, with 0.1 M NaCl, 1mM EDTA, and 5% [w/v] TRITON X-100).
4. Add 25 μl of a freshly prepared lysozyme solution (10 mg/ml in 10 mM Tris-HCl, pH 8.0).
5. Mix by vortexing for 3 seconds.
6. Incubate the lysis mixture for 30 minutes at 37 °C
7. After incubation, place the tube containing the lysis mixture in a boiling water bath for exactly 40 seconds.
8. Centrifuge the lysis mixture at 14,000 g.
9. Remove the pellet (cell debris) from the tube using a sterile toothpick.
10.Plasmid DNA from the supernatant may then be purified and analyzed.
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