1. Grow an overnight culture from a single colony in 2 mls LB + antibiotic

2. Transfer 1.5 mls of culture to eppendorf tube. Spin for 1 minute on high and remove supernatant.

3. Add 700 µl STET. Add 25 µl lysozyme stock solution. Vortex to resuspend pellet. Place tubes on ice for 5-10 minutes.

4. Boil tubes for 1 minute.

5. Spin in microfuge for 10 minutes.

6. Pull out snot pellet with a toothpick.

7. Add 700 µl of room temperature isopropanol. Mix tubes by inversion.

8. Spin in a microfuge for 10 minutes. A small white/clear pellet will form.

9. Remove supernatant. Wash with 70% ethanol.

10. Dry pellet and resuspend in 50µl water containing 20 µg/ml RNase A.

STET

8% sucrose 5% Triton X-100 50 mM Tris-HCl, pH 8.0 50 mM EDTA, pH 8.0

store at 4°C

Lysozyme Stock Solution (10 mg/ml)

150 mg lysozyme 3.75 mls 1M Tris-HCl, pH 8.0 11.25 mls H2O

store at -20°C.