1. Thaw the competent E.coli (DH5alpha) cells on ice (not in hands!)
2. As soon as it is thawed, add 50ul of the cells to the DNA (~50-100ng of pure plasmid)
3. Let it rest on ice for 20-30 min. Meanwhile, put agar plate (with correct antibiotic)

at 37°C for prewarming.

1. Put the tube with DNA+E.coli at 42°C for 45 sec-1 min (heat shock)
2. Add 400ul of LB broth and place at 37°C for 20-30min (shaking)
3. Spread the cells on the prewarmed plate (and let it dry)
4. Incubate the plate upside-down at 37°C for ~14-15h (leaving it more than 16h decreases

the plasmid quality)