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Team:UEA-JIC Norwich/ligation
From 2011.igem.org
UNIVERSITY OF EAST ANGLIA-JOHN INNES CENTRE
1. Add 2µl of 10× T4 DNA ligase buffer to an eppendorf tube.
2. Add 50ng vector DNA to the mix.
3. Add 50ng insert DNA to the mix.
4. Add 20µl nuclease free water to the tube.
5. Add 1µl of T4 DNA ligase to the tube.
6. Gently mix the reaction via pipetting up and down.
7. Incubate at room temperature for 10 minutes to acquire sticky ends.
8. For Blunt ends incubate at room temperature for 2 hours.
9. Chill on ice and then use in transformation.
