FAQ: How much DNA should be used in a ligation using T4 DNA Ligase?

The unit definition uses 0.12 μM (300 μg/ml) lambda HindIII fragments. The high DNA concentration can be used for linker ligation. To promote circle formation, useful in transformation, a lower total DNA concentration should be used. The overall concentration of vector + insert should be between 1-10 μg/ml for efficient ligation. Vector:Insert molar ratios between 1:1 and 1:10 are recommended (1:3 is typical). If you are unsure of your DNA concentrations, perform multiple ligations with varying ratios.
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    Quick Tips - How do I calculate how much DNA to add to a ligation reaction?

    This video provides tips for calculating the ideal insert:vector molar ratio before performing a ligation reaction.