9°NDNALigaseisnotasubstituteforT4 DNALigase.Thecohesiveendunitisequivalenttothenick-closingunitofBaranyetal.IncubateDNAandenzymein1X9°NDNALigaseReactionBufferat45°Cfor15minutesorinathermocyclerwithaprogramsuitedtothereactiondescribedbyBarany(1991)GeneticDiseaseDetectionandDNAAmplificationUsingClonedThermostableLigase.Proc.Natl.Acad.Sci.USA88,189-193.Thereactionisstoppedwithamixtureof50%glycerol,50 mM EDTA,bromphenolblue.