Team:Tuebingen/Notebook/Protocols/ligation

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Ligation

Reagents

20 - 50 ng Linearized vector
1 µL 10x T4 DNA Ligase Buffer
1 µL T4 DNA 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. Vortex and incubate at 4 °C over night. On the next day, inactivate enzymes at 70 °C for 5 min. Store at -20 °C.

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