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T4 DNA Ligase 2u. Vortex the tube and spin down in a microcentrifuge for 3-5 seconds.
Incubate the mixture for 1 hour at 22°C .
Inactivate T4 DNA Ligase by heating the reaction mixture at 65°C for 10 minutes. The resulting ligation products can be digested directly with restriction endonucleases.
Note
Some restriction endonucleases are susceptible to star activity in ligation buffer. Therefore, DNA should be precipitated with isopropanol or ethanol after ligation and dissolved in an appropriate buffer.