Home Feedback Search Online Order CATALOG

International 

+32 (0) 16 58 90 45

+32 (0) 16 50 90 45

France

01 43 25 01 50

01 43 25 01 60

Italy

02 36 00 65 93

02 36 00 65 94

Germany

0241 6085 13140

0241 6085 33033

Back Up Next


 

 

T4 DNA Ligase datasheet

 

T4 DNA ligase
(Cat. No.: E003)

Description:
    T4 DNA Ligase is a recombinant enzyme formulated and tested for high activity in blunt- and cohesive-end ligations for cloning and linker addition. The enzyme covalently joins 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA. It is free of detectable exo- or endonuclease activities.

Source:
    Purified from recombinant E. coli strain.

Applications:
    Cloning of restriction fragments.
    Joining linkers and adapters to blunt-ended DNA.

Unit Definition:
    One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of  lambda DNA (5' DNA termini concentration of 0.12M, 300g/ml) in a total reaction volume of 20l in 30 minutes at 16oC in 1X T4 DNA Ligase Reaction Buffer.

Concentration:
    400,000 units/ml

Quality Control Tests:
    Activity, SDS-PAGE (purity), DNase, endonuclease/nickase, RNase.

Storage:
    T4 DNA ligase in 10 mM Tris-HCl, 50mM KCl, 1mM dithiothreitol, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol, pH 7.4 (25oC) should be stored at -20oC.

Heat Inactivation:
    65oC for 10 minutes

1X T4 DNA Ligase Reaction Buffer:
    50mM Tris-HCl, 10mM MgCl2, 1mM ATP, 10mM dithiothreitol, 25g/ml BSA, pH 7.5 (25oC)

Reaction Conditions:
    1X T4 DNA Ligase Reaction Buffer, incubate at 16oC.
    For convenience, ligations may be done at room temperature (20-25oC). For cohesive (sticky) ends, use 1l of T4 DNA Ligase in a 20l reaction for 10 minutes. For blunt ends, use 1l of T4 DNA Ligase in a 20l reaction for 2 hours.

References:
    Engler, M.J. et al. (1982) P.D. Boyer (Eds.), The Enzymes, 5, pp. 3. San Diego: Academic Press.
    Remaut, E. et al. (1983) Gene, 22, 103-113.
    Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, (2nd Ed.), 1.53-1.73.
    Weiss, B. et al. (1968) J. Biol. Chem., 243, 4543-4555.
 

 

 

For more information :: Bioxys and Gentaur BVBA :: Av. de l'Arme 68 B4 :: BE-1040 BELGIUM

Email: info@gentaur.com
 

International 

+32 (0) 16 58 90 45

+32 (0) 16 50 90 45

France

01 43 25 01 50

01 43 25 01 60

Italy

02 36 00 65 93

02 36 00 65 94

Germany

0241 6085 13140

0241 6085 33033

Send mail to webmaster with questions or comments about this web site.
Copyright 2005 Gentaur BVBA
Last modified: feb-07