Template:Team:Groningen/CONTENT/PROTOCOLS/Ligation

Ligation

Prepare the following reaction mixture.

Ingredient

Amount

Linear vector DNA

20 – 100 ng

10x T4 DNA ligase buffer

2 μl

T4 DNA ligase

1 U

Water, nuclease-free (= sterile milliQ)

to 20 μl

Total volume

20 μl

Ligation mix.

Incubation

Incubate for 10-60 minutes at 22°C.

Transformation

Use up to 5 μl of the mixture for transformation of 50 μl chemically competent cells or 1-2 μl per 50 μl electrocompetent cells.