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Ligation

Ligation means literally "gluing together." In molecular biology, DNA ligation involves linking together two strands of DNA that each have double-strand breaks –- a break in each of the complementary strands. (Single-strand breaks are easily fixed with DNA polymerase by using the opposite strand as a template.)

Ligation is a natural process taking place in both DNA repair and DNA replication, but it has been adopted by genetic engineering laboratories for its usefulness in recombinant DNA experiments and applications. For instance, through using ligation, experimenters can mix cut plasmids and free-floating genes with complementary "sticky" ends, and then introduce a ligase; its catalytic action will attach the gene and the plasmid. The most vital part of this operation is to maintain the experiment at an optimal temperature, which differs from case to case. High temperatures disrupt hydrogen bonding, while low temperatures prevent catalysis from taking place. Short fragments require a much lower temperature over a much longer time.

DNA ligases used in ligation were originally found in E. coli and other bacteria.

Web Resources On Ligation

DNA Ligation
DNA Ligation Protocol

Book Resources On Ligation

From Genes to Genomes : Concepts and Applications of DNA Technology by Dale & von Schantz
DNA Science: A First Course by Micklos et. al.

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