Short explanation:

DNA ligation is necessary to assemble digested DNA parts into a vector. The cut ends generated by restriciton enzymes are put together by DNA ligase.

Procedure: 20 µL batch

- T4 Ligase Buffer 2 µL

- T4 Ligase 1 µL

- digested vectors 25 ng (Molar ratio = Insert : backbone 3:1)

- add ddH2O to 20µL

- Incubate at 16°C overnight, alternatively incubate at room temperature for 30 min (results might be worse)

- Inactivate 20min at 80°C