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T4 DNA Ligase Recombinant ( T4 DNA )
DescriptionT4 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
Data sheet
| Formulation | 50mM Tris-HCl (pH 7.8 at 25°C), 10mM MgCl2, 10mM DTT, 1mM ATP, 25 ug/ml BSA and DNA (0.1 to 1 um in 5 termini). Optimal ligation occurs at 16C. |
| Applications | Cloning of restriction fragments.Joining linkers and adapters to blunt-ended DNA. |
| 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. |
| Expression host | Escherichia Colilambda lysogen NM 989. |
| Synonyms | DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. |
| Reagent Appearance | Sterile filtered liquid formulation having a concentration of 167,000 U/ml. |
| Biological Activity | One Weiss unit is equivalent to circa 67 cohesive-end ligation units. T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM. Ligation of blunt-ended and single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 (10% w/v final concentration) or hexamine chloride, or by reducing the ATP concentration to 50uM. To dilute T4 DNA Ligase that will subsequently be stored at 20°C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. |
| Unit Defenition | 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of DNA (5 DNA termini concentration of 0.12 uM, 300- ug/ml) in a total reaction volume of 20 ul in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer. 2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. |