For details on NEB's quality controls for DNA ligases, visit our Ligase Quality page.
T4 DNA Ligase Competitor Study - Nuclease Contamination
T4 DNA Ligase from multiple suppliers was tested in reactions containing a fluorescent labeled single stranded, double stranded blunt, 3’overhang or 5’ overhang containing oligonucleotides. The percent degradation by contaminating nucleases is determined by capillary electrophoresis and peak analysis. The resolution is at the single nucleotide level.
Highlights
Isolated from a recombinant source
Supplied with 10X Reaction Buffer
Product Source
An E. coli strain that carries the cloned T4 DNA Ligase gene.
This product is related to the following categories:
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5´ DNA termini concentration of 0.12 µM, 300- µg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
Concentration: 400,000 cohesive end units/ml and 2,000,000 cohesive end units/ml
Reaction Conditions
1X T4 DNA Ligase Reaction Buffer
Incubate at 16°C
1X T4 DNA Ligase Reaction Buffer
50 mM Tris-HCl
10 mM MgCl2
1 mM ATP
10 mM DTT
(pH 7.5 @ 25°C)
Storage Buffer
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Sexton T, Kurukuti S, Mitchell JA, Umlauf D, Nagano T, Fraser P (2012) Sensitive detection of chromatin coassociations using enhanced chromosome conformation capture on chip Nat Protoc; 7(7), 1335-50. PubMedID: 22722369, DOI: 10.1038/nprot.2012.071
Publications
Thuronyi, B.W., Koblan, L.W., Levy, J.M. et al (2019) Continuous evolution of base editors with expanded target compatibility and improved activity Nat Biotechnol; 37, 1070-1079.. DOI: 10.1038/s41587-019-0193-0
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