DNA ligation
General Information
DNA ligation is a commonly used protocol in molecular biology. DNA ligase is used to create a phophodiester bond between the 5' phosphate and 3' hydroxyl groups of DNA. Most commonly, one needs to insert a DNA sequence of interest into a plasmid. Ideally, you can cut the DNA and vector with the same restriction sites. Then you can add both DNA sequences, the cut insert and vector sequence, and ligate them together to form circular vector using DNA ligase. T4 DNA ligase is the most commonly used DNA ligase for molecular biology techniques.
General Procedure
The prodedure for ligation depends upon whether the ligation is a blunt-end ligation or a ligation involving overlaps. Generally, you add the DNA you want to ligate, and then add a DNA ligase to ligate them together.
