| 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.
Applications:
- cloning of restriction fragments;
- joining linkers and adapters to blunt-ended DNA
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| 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.) |
| Optimal temperature | 16oC |
| 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.
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| Quality control | The enzyme is purified free of contaminating endonucleases and
exonucleases.
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| MSDS: | Download MSDS as PDF |
