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
