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Some engineering has long been done to improve the in vitro activity of T4 DNA ligase; 1 productive technique, one example is, examined T4 DNA ligase fused to many choice DNA binding proteins and located the constructs with both p50 or NF-kB as fusion partners have been around a hundred and sixty% extra Lively in blunt-conclude ligations for cloning uses than wild type T4 DNA ligase.[six] A standard reaction for inserting a fragment right into a plasmid vector would use about 0.01 (sticky finishes) to 1 (blunt ends) units of ligase. The optimal incubation temperature for T4 DNA ligase is 16 °C.[citation necessary]
DNA ligase plays an important function in DNA replication, DNA repair service, and DNA recombination. Now day purified DNA ligase is isolated during the laboratory, and that is used get more info in gene cloning to hitch DNA molecules with each other to form recombinant DNA.
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It is more steady and Lively at Severe temperatures than common DNA ligases. It's used in DNA amplification reactions to detect mutations in mammalian DNA.
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