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Ligation Calculator

Chemistry & Biology

Calculate the mass of DNA insert needed for a cloning ligation reaction at a target insert-to-vector molar ratio.

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ng
0.001 ng1,000,000 ng
bp
1 bp1,000,000,000 bp
bp
1 bp1,000,000,000 bp
0.0011,000

Required Insert Mass

25.000

The mass of insert DNA needed to achieve the target molar ratio in the ligation reaction.

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Odeh Ahwal

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Odeh Ahwal
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Frequently Asked Questions

What is a ligation reaction?

A ligation reaction joins a DNA insert fragment into a vector, such as a plasmid, using an enzyme called DNA ligase, a standard step in molecular cloning.

Why does the molar ratio matter?

DNA is measured by mass, but a ligation reaction depends on the molar amount of insert relative to vector, since it is individual DNA molecules that pair up. Insert and vector fragments of very different lengths need different masses to reach the same molar ratio, so the mass-to-length relationship has to be converted through the molar ratio.

What is a typical insert:vector molar ratio?

A 3:1 insert-to-vector molar ratio is a commonly used starting point for many cloning protocols, though the ideal ratio can vary by protocol and is often optimized experimentally.

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