The following protocol is designed for NEB 10-beta Competent E. coli (Cloning Efficiency) provided in this kit. Important – please read the FAQs regarding competent cell requirements before using a different cell strain.
Note: For further general discussion about transformations with NEB 10-beta Competent E. coli, please refer to the transformation protocol on the C3019 product page. The cloning efficiency cells provided with this kit differ from the high-efficiency strain in their transformation efficiencies only.
- 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 20–30 minutes.
- Heat shock at 42°C for 30 seconds.
- Chill on ice for 5 minutes.
- Add 950 μl of SOC and place at 37°C for 60 minutes with rotation or shaking at 250 rpm (outgrowth step).