E. coli (DH5α chemically-competent) Transformation

1. Thaw one aliquot of DH5α competent cells (from –80°C freezer) on ice
2. Add 5 μl DNA and 100 μl competent cells to new cell culture tube
3. Incubate on ice for 30 minutes
4. Heat shock in 42°C water bath for 30 seconds
5. Cool on ice
6. Add 900 μl liquid media (e.g. LB) to cell culture tube
7. Shake at 37°C for 40-60 minutes
8. Centrifuge at 5000 rpm for two minutes
9. Remove all but 100-150 μl of the LB and resuspend pellet
10. Spread 100-150 μl on plate of media + antibiotic (e.g. LB agar + carb)
11. Incubate plates up-side-down, overnight at 37°C