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DNA ligation: Difference between revisions
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Barry Canton (talk | contribs) |
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This is a [[Help:Consensus protocol|consensus protocol]] see the bottom of this article for specific protocols. | This is a [[Help:Consensus protocol|consensus protocol]] see the bottom of this article for specific protocols. | ||
DNA ligation is the process of joining together two DNA molecules | DNA ligation is the process of joining together two DNA molecule ends (either from the same or different molecules). Specifically, it involves creating a [[Wikipedia:phosphodiester bond|phosphodiester bond]] bond between the 3' hydroxyl of one nucleotide and the 5' phosphate of another. This reaction is usually catalyzed by a [[Wikipedia:DNA ligase|DNA ligase enzyme]]. This enzyme will ligate DNA fragments having blunt or overhanging, complementary, 'sticky' ends. Typically, it is easier to ligate molecules with complementary sticky ends than blunt ends. [[T4 DNA ligase]] is the most commonly used [[Wikipedia:DNA ligase|DNA ligase]] for molecular biology techniques and can ligate 'sticky' or blunt ends. | ||
Most commonly, one needs to insert a DNA molecule of interest into a plasmid, ready for [[transformation]] into [[competent cells]]. Ideally, DNA and vector are individually cut with the same restriction enzyme, then both are added to a ligation reaction to be circularised by DNA ligase. | Most commonly, one needs to insert a DNA molecule of interest into a plasmid, ready for [[transformation]] into [[competent cells]]. Ideally, DNA and vector are individually cut with the same restriction enzyme, then both are added to a ligation reaction to be circularised by DNA ligase. | ||
