Team:Tuebingen/Notebook/Protocols/ligation
From 2013.igem.org
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 |
to 10 µL | Aqua dest. |
Procedure
Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).