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From 2010.igem.org

Transformation

1. 1:3
2. 1:5
3. Vector (no insert), negative control for ligation
4. DH5Alpha (without plasmid)
5. DH5Alpha (with plasmid, pSB1AT3)

Protocol:

1. Thaw a 200µl aliquot of E. coli DH5Alpha. Add the transforming DNA. We added 5µl of vectors from tubes 1 to 3 and 1µl of vector from tube 5 because vector from tube 5 is has not been ligated whereas tubes 1 to 3 have been ligated and will contain cells in different forms.
2. Incubate for 45 min on ice.
3. Heat-shock the cells at 42°C for 120 secs, and place on ice again for 3-4 min.
4. Add 1ml of LB broth to each tube and incubate the cells at 37°C for 1.5 hr (static for initial 20 mins, shaking for remaining time).
5. Plate out 200 µl/plate on LB (agar at 1.5%), with ampicilin.
6. Incubate plates overnight at 37°C.