STETL DNA Miniprep

1. Grow 2 mL bacterial culture overnight.
2. Pulse spin 1.5 mLs of culture in Eppendorf tube.
3. Pour off supernatant. Resuspend pellet in remaining liquid.
4. Add 300 µL STETL. Vortex well.
5. Place in 100°C heat block for 90 seconds.
6. Spin 10 minutes in room temperature microfuge.
7. Pour top layer into new Eppendorf tube containing 300 µL isopropanol. Vortex well.
8. Spin 5 minutes in room temperature microfuge. Drain supernatant.  Air dry tubes for >5 minutes.
9. Resuspend in 100 µL ddH2O. Use 5 µL for normal restriction digest.

STETL Recipe		To make 100 mLs
8% Sucrose 5% Triton X100 50 mM EDTA, pH 8.0 50 mM Tris, pH 8.0 1 mg/mL lysozyme water to final volume		8 gm Sucrose 5 mLs Triton X100 10 mLs 0.5 M EDTA, pH 8.0 5 mLs 1.0 M Tris, pH 8.0 100 mg lysozyme final volume = 100 mLs 10 mL aliquots into 15 mL Falcon tubes; store at -20C.