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Applications	Substrate	Reaction catalyzed	Inactivation	Ligase
Joining dsDNA with cohesive or blunt termini Joining of ds oligonucleotide linkers or adaptors to dsDNA Repairing nicks in duplex DNA, RNA or DNA-RNA hybrids Ligase-mediated RNA detection Site-directed mutagenesis Amplified fragment length polymorphism	dsDNA dsRNA dsDNA-RNA hybrids ds poly- and oligonucleotides	ATP dependent intra- and intermolecular formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.	Heating at 65°C, 10 min or 70°C, 5 min	T4 DNA Ligase
RNA 3'-end labeling with cytidine 3',5'-bis [alpha-³²P] phosphate Joining RNA to RNA Circularization of synthetic oligonucleotides Specific modifications of tRNAs Oligodeoxyribonucleotide ligation to single-stranded cDNAs for 5'-RACE Site-specific generation of composite primers for PCR	ssDNA ssRNA poly- and oligonucleotides	ATP dependent intra- and intermolecular formation of phosphodiester bonds between 5'-phosphate and 3'-hydroxyl termini of poly- and oligonucleotides, ssRNA and ssDNA	Heating at 70°C, 10 min	T4 RNA Ligase

ss – single-stranded
ds – double-stranded