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Products >  Molecular_Biology >  Modifying_Enzymes >  Ligases >  T4_DNA_Ligase

T4 DNA Ligase

T4 DNA Ligase catalyzes the formation of phosphodiester bonds between juxtaposed 5' phosphate and 3' hydroxyl termini in double-stranded DNA using ATP as a coenzyme. Both blunt and cohesive end DNA ligation, as well as single-stranded nick repair of DNA, RNA and DNA/RNA, are possible via the T4 DNA ligase. This enzyme is supplied in a buffer of 10 mM Tris-HCl (pH 7.5), 50 mM KCl, 10 mM 2-mercaptoethanol, 0.1 mM EDTA and 50% glycerol.

At-A-Glance Documents Images & Data Resources

Applications

  • Blunt and cohesive end DNA ligation
  • Blunt end linker addition
  • Single-strand DNA, RNA and DNA/RNA repair
  • DNA fragment insertion into vectors

Components

2011A: 200 Weiss units
2011B: 5 x 2011A (1000 Weiss units)

Source

Recombinant E. coli

Storage

–20°C

Unit Definition

One ligation unit is defined as the amount of enzyme that ligates >90% of 6 µg of Hind III-restricted lambda DNA in a 20 µL reaction volume in 30 minutes at 16°C. One unit of the enzyme corresponds to 0.008 Weiss units by the ATP-PPi exchange reaction in 66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl2, 10 mM DTT, 66 µM ATP and 3.3 µM [32P]Na4P2O7.

References

1. Weiss, B. et al. (1968) J. Biol. Chem. 243:4543-4555.

2. Weiss, B. et al. (1968) J. Biol. Chem. 243:4556-4563.

Buffer

Supplied with 10X Reaction Buffer [660 mM Tris-HCl (pH 7.6), 66 mM MgCl2, 100 mM DTT and 1 mM ATP].

Concentration

350 U/µL (2.8 Weiss units/µL)




 
 
Products
Cat. # Product Package Size Price # of Units Select
2011A T4 DNA Ligase 25,000 Units $45.00
2011B T4 DNA Ligase 125,000 Units $180.00
 

 


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