4 DNA ligase uses the energy of ATP to catalyze the formation of phosphate from the 3´ hydroxyl end and the 5´ phosphate end between double-stranded DNA. diester bond. This unique T4 DNA ligase buffer optimizes ligation and can be completed in 5 minutes (1). Single-stranded nucleic acids are not substrates for this enzyme. Provides AT4 DNA Ligase technical bulletin.

Applications: Cloning (blunt or sticky end ligation) (2). Add adapters to blunt-ended DNA (2).

Source: Purified from colilysinœ original NM989.

Performance and quality testing: Endodeoxyribonuclease, 3´ and 5´ exodeoxyribonuclease assays; tested for ligation efficiency.

Unit definition: One unit catalyzes the exchange of 1 nmol ³²P-labeled pyrophosphate to ATP in 20 minutes at 37°C. (One unit is approximately equal to 300 sticky end ligation units)

Unit reaction conditions:66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl₂ , 10 mM DTT, 66 μM ATP, 3.3 μM 32 P-labeled pyrophosphate and enzyme, 0.1 ml, 20 min, 37°C.