Team:Baltimore/Protocols/transformation

From 2011.igem.org

Bacterial Transformation Protocol

1. Prepare water bath @ 42˚C.
2. Obtain and label LB/Amp plates
3. Obtain SOC media, check for sterility
4. Transfer 1µl ligation mix into 1.7ml eppendorf tubes. Place on ice; tubes must be at 0˚C before adding cells. Aliqout 25µl of competent cells per tub.
5. Incubate on ice for 30 min.
6. Heat shock @ 42˚C for (exactly!) 45 seconds.
7. Immediately place heat shocked cells back on ice for 2 min.
8. Add 400µl SOC and incubate in 37˚C incubator for 30 min.
9. Pipette entire mixture onto LB/Amp plates and spread.
10. Incubate plates overnight at 37˚C. (Store upside down to prevent condensation; incubate for no more than 18hrs before parafilm wrapping and storing @ 4˚C.