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From 2010.igem.org

Reagents

• T4 DNA ligase
• 10x T4 DNA Ligase Buffer
• Deionized, sterile H2O
• Purified, linearized vector (likely in H2O or EB)
• Purified, linearized insert (likely in H2O or EB)

Equipment

• Vortex

Procedure
10μL Ligation Mix
Note: Larger ligation mixes are also commonly used

• 1.0 μL 10X T4 ligase buffer
• 6:1 molar ratio of insert to vector (~10ng vector)
• Add (8.5 - vector and insert volume)μl ddH2O
• 0.5 μL T4 Ligase