(Quality is sufficient for RE Digestion and for sequenzing reaction, outcome is around the double of a normal alkaline lysis)

DAY 1:
Inoculate 3-5ml of LB conaining the correct antibiotic (Amp for overnight liquid cultures is usually 100mg/ml) from a single colony, and incubate overnight.
 

DAY2:
1.    Transfer 1.5ml of the culture to an eppendorf tube and spin down for 10'' at maximum speed. Remove the supernatant  by aspiration and leave the tubes inverted on a
        kimwipe for  sometime.
2.    Resuspend the bacterial pellet in 350ml  STET-solution. Be sure to resuspend  completly.
3.    Add 25ml of a 10mg/ml Lysozym in 10mM TrisCl, ph 7.5. Vortex and led stand on ice for five minutes.
4.    Boil the mixture for a short while, I usually do it for 90''. Immediatly transfer the tubes to the centrifuge and centrifuge for 15-20'.
5.    Take out the white gummy pellet (sometimes it has a fluffy appearance in which case you had some infection. No need to continue) with a sterile toothpick.
6.     Add 40 ml Sodiumacetate (2,5M, pH 5,2) and 420 ml Isopropanol, mix by vortexing and centrifuge for 20' at 4^oC. A huge white pellet wiil appear, no worry, this is normal.
7.    Wash the pellet with 70% Ethanol ( -20^oC is better, but roomteperature will do).
8.    Resuspend the pellet in 50ml TE suplemented with 10mg/ml DNAsefree RNAse A. Have Fun doing whatever you want with this DNA, if you want to use it directly  in
       delicate things, I would suggest a Phenol/CHCL₃ extraction and than it should perform pretty well in any application tou can think of.
 

            STET Solution:
                    -0.1M NaCl
                    -10mM TrisCl (pH 8.0)
                    -1mM EDTA (pH 8.0)
                    -5% Triton X-100 (v/v)