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AAT Bioquest

What is blunt end ligation?

Posted November 4, 2020


Answer

Ligation refers to the joining or ligating of two nucleic acid fragments through the action of an enzyme. The attaching of blunt-ended DNA fragments by the enzyme DNA ligase is known as blunt end ligation.

This is a crucial laboratory procedure used in the molecular cloning of DNA. During the process the linearized plasmid vector and the blunt-ended insert are mixed with DNA ligase. The reaction gets initiated when the vector, insert and ligase come together in solution. A major advantage of blunt-end cloning is that the desired insert does not require any restriction sites in its sequence as blunt-ends are usually generated in a PCR, and the PCR generated blunt-ended DNA fragment may then be ligated into a blunt-ended vector generated from restriction digest.

Additional resources

A novel series of high-efficiency vectors for TA cloning and blunt-end cloning of PCR products

Portelite™ Fluorimetric High Sensitivity DNA Quantitation Kit *Optimized for CytoCite™ and Qubit™ Fluorometers*