|Source||Isolated from E.coli strain that carries the cloned DNA ligase gene from bacteriophage T4|
|Description||T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5` phosphate and 3` hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt-end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA, or DNA/RNA hybrids.|
- cloning of restriction fragments;
- joining linkers and adapters to blunt-ended DNA
|Unit definition||One unit is defined as the amoumt of enzyme required to give 50% ligation of Hind III fragments of Lambda DNA (5` DNA termini concentration of 0.12 μM [300 μg/ml]) in 20 μl of 1 x T4 DNA Ligase Reaction Buffer in 30 minutes at 16°C. ATP is an essential cofactor for the reaction
|Reaction buffer||SE-buffer DNA ligase T4, (50 mM Tris-HCl (pH 7.8 at 25°C); 10 mM MgCl2; 10 mM DTT; 1 mM ATP.)|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.|
|Notes||A precipitate may be observed in T4 DNA Ligase Buffer after defrosting.|
Before the first use we recommend to heat this buffer at 37°C for 10-15 min and dissolve the precipitate by shaking.
Also we recommend to dispense the buffer into small aliquots and store them at -20°C.
Avoid defrosting the buffer more than 2-3 times. The aliquot of T4 DNA Ligase Buffer may be stored at +4°C during 7 days.
|Quality control||The enzyme is purified free of contaminating endonucleases and
|MSDS:||Download MSDS as PDF|