Team:Tuebingen/Notebook/Protocols/ligation

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Ligation
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Reagents

20 - 50 ng Linearized vector
1 µL 10x Ligase buffer
1 µL T4 Ligase
5 µl
(up to 5:1 molar ratio insert to vector)
Insert DNA
fill up to 10 µL Aqua dest.

 

Procedure

Mix all reagents and fill up with Aqua dest. to 10 µL and incubate at 4 °C over night. On the next day, inactivate enzymes at 70 °C for 5 min.