Transformation Protocol for NEB PCR Cloning Kit
- Thaw a 50 μl tube of competent cells on ice for 10 minutes.
- Add 2 μl of ligation reaction; gently mix by flicking the tube 4–5 times.
- Incubate on ice for 20 minutes.
- Heat shock at 42°C for 30 seconds.
- Chill on ice for 5 minutes.
- Add 950 μl of NEB™ 10-beta/Stable Outgrowth Medium and place at 37°C for 60 minutes with rotation or shaking at 250 rpm (outgrowth step).