NxGen T4 DNA Ligase
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The low concentration ligase includes NxGen T4 DNA Ligase at 2 U/µL and 10X T4 DNA ligase Buffer. The high concentration ligase includes NxGen T4 DNA Ligase at 10 U/µL and both 2X Rapid Ligation Buffer and 10X T4 DNA ligase Buffer.
NxGen T4 DNA Ligase
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- Reliably join double-stranded nucleic acid chains.
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T4 DNA Ligase catalyses 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||
|2 U/µL||10X T4 DNA Ligase Buffer|
|High Concentration Rapid Kit||1,500 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 MgCl 2, 50 mM dithiothreitol, 10 mM ATP, pH 7.6 @ 25 °C.
2X Rapid Ligation Buffer is composed of 132 mM Tris-HCI, 20 mM MgCl 2, 2 mM dithiothreitol, 2 mM ATP, 15% PEG, pH 7.6 @ 25 °C.
Figure 1: Purity
The purity of the T4 DNA ligase is equal or greater than 95% as judged by SDS-polyacrylamide gel electrophoresis with Coomassie blue staining. Molecular weight markers are in Lane 1. T4 DNA ligase was loaded at 5 units (Lane 2) and 10 units (Lane 3).
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.
Storage: -20 °C
Source: A recombinant E. coli strain carrying the cloned T4 DNA Ligase gene.