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From 2013.igem.org

Extract the target DNA.
Materials: bacteria(in the liquid culture), reagent case.

Methods:

1. After centrifuging the bacteria, collect them in the same EP tube for many times, then leave the supernate out. 12100r/min every time, and last for 1min.

2. Inject 250ul S1 buffer into the bacteria sediment, then make them in suspension.
3. inject 250ul S2 buffer(contain RNA polymerase)(alkaline), put the EP tube upside down to split them.
4. Inject 350ul S3 buffer(acidic), put the EP tube upside down gently, then centrifuge them for 10 mins.(the time break between process 3 and 4 cannot be longer than 5 mins.)
5. Absorb the supernate and inject them into the pilar, then centrifuge for 1min and desert the waste.
6. Inject 500ul W1 buffer, then centrifuge for 1min.
7. Inject 700ul W2 buffer, then centrifuge for 1min.

8. Change the waste tube into EP tube. Then inject 30ul ddH2O into the pilar, then stew it and centrifuge for 1min.Eventually put it into the -80’c refrigerator.