Invitrogen E. coli DNA Ligase
Catalyzes formation of phosphodiester bonds in presence of β-NAD between double-stranded DNAs with 3' hydroxyl and 5' phosphate cohesive termini
Manufacturer: Invitrogen 18052019
Single-stranded nucleic acids are not substrates for this enzyme.
- Source: Purified from E. coli 594 (Su-) bearing λ lysogen gt4lop-11 lig+ S7
- Performance and Quality Testing: Endodeoxyribonuclease and 3' and 5' exodeoxyribonuclease assays; ligation efficiency tested
- Unit Definition: One unit is amount of enzyme required to give 50% ligation of Hind III-digested λ DNA in 30 min. at 16°C in a final volume of 20µL containing a 5' termini concentration of 0.12 'M (300µg/mL)
- Unit Reaction Conditions: 18.8mM Tris-HCl (pH 8.3), 90.6mM KCl, 4.6mM MgCl2 , 3.8mM DTT, 0.15mM λ-NAD, 10mM (NH4 )2 SO4 in 20µL for 1 hr. at 16°C
Second-strand cDNA synthesis; T4 DNA Ligase alternative when blunt-end ligation is not required; Cloning, Restriction Enzyme Cloning
Shipping Conditions: Approved for shipment on wet or dry ice
|10X Reaction Buffer [188mM Tris-HCl (pH 8.3), 906mM KCl, 46mM MgCl2 , 37.5mM DTT, 1.5mM λ-NAD, 100mM (NH4 )2 SO4 ]|
|Store at -20°C|
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For Research Use Only. Not for use in diagnostic procedures.