E. Coli mini plasmid prep.

1. Transfer 1-1.5 mls of the culture into an eppendorf tube.
2. Spin 1 min. at 15K room temp. (remove with vacuum pump siphon)
3. Resuspend in 300 µls of STET containing 1mg/ml lysozyme.
4. Vortex, then Boil for 1 min. in the 100C temp block.
5. Spin 10 min. at 15 K room temp.
6. Remove and discard pellet using a tooth pick.
7. Add 300 µls of cold Isopropanol, mix and leave at -20^oC for 30 min. to 1 hour.
8. Spin 15 min. at 15K 4^oC. (remove supernatant with vacuum pump siphon)
9. Wash with 70% EtOH. (remove supernatant with vacuum pump siphon)
10. Dry pellets for 10 min. r/t. and resuspend in 50 µls of 1xTE containing 50µg/ml RNase.
11. Incubate for 10 min. at 37^oC.
12. Analyse on agarose gel.
    

1x STET	100mls
8% Sucrose	8 gms Sucrose
50 mM Tris pH8.0	5 mls 1M Tris pH 8.0
50 mM EDTApH8.0	10 mls 0.5 M EDTA pH8.0
5% Triton X-100	5 mls Triton X-100
	H2O to 100mls

for 18 samples prepare 6 mls of STET sol'n w/ 6 mgs of lysozyme

1XTE w/ 50µg/ml RNase : add 5µls of a 10 mg/ml RNase stock to 1ml 1XTE