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- Reliably join double-stranded nucleic acid chains.
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the terminal 5′ phosphate and 3′ hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
| T4 DNA Ligase Version | Sizes | Ligase Concentration | Provided Buffer |
| Low Concentration Ligase |
1,500 U |
2 U/µL | 10X T4 DNA Ligase Buffer |
| High Concentration Rapid Kit | 1,500 U 7,500 U |
10 U/µL | 2X Rapid Ligation Buffer, 10X T4 DNA Ligase Buffer |
10X T4 DNA Ligase Buffer is composed of 500 mM Tris-HCI, 100 mM MgCl2, 50 mM dithiothreitol, 10 mM ATP, pH 7.6 @ 25 °C.
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Figure 1: Purity |
2X Rapid Ligation Buffer is composed of 132 mM Tris-HCI, 20 mM MgCl2, 2 mM dithiothreitol, 2 mM ATP, 15% PEG, pH 7.6 @ 25 °C.
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Figure 2: Exonuclease and Endonuclease Activity Assay The T4 DNA ligase is free of detectable of exo- and endonuclease activities, as judged by agarose gel electrophoresis following incubation of 10 units of enzyme for 16 hours at 37 °C with 1 µg of HindIII-digested λ DNA (Lanes 3 and 4) and supercoiled pUC19 DNA (Lanes 5 and 6). Molecular weight markers are in Lanes 1 and 2. Unit Definition: One Weiss Unit is defined as the amount of enzyme required to convert 1 nmol of 32P-labeled inorganic pyrophosphate into Norit adsorbable material in 20 minutes at 37 °C, using specified reaction conditions. Note: 1 Weiss Unit is approximately 67 cohesive end units. Source: A recombinant E. coli strain carrying the cloned T4 DNA Ligase gene. |
