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T4 DNA Ligase

Looking for additional T4 DNA Ligase formulations and variants? Learn More

This product is available in a glycerol-free format. Contact us for more information.
GMP-grade reagent also available. Learn more.

T4 DNA Ligase will ligate these substrates:

dsDNA

Nicked DNA/RNA

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 and some DNA/RNA hybrids (1).
The industry standard for performance and quality
Applicable for use in high-complexity library cloning
This product can be purchased in larger volumes. Submit an inquiry to find out more.
Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart or DNA Ligase Selection Chart

Catalog #	Cat #	Concentration	Conc	Size	List Price
M0202S	400,000 units/ml	20,000 units	$71.00	$71.00	Sign In Sign In
M0202T	2,000,000 units/ml	20,000 units	$71.00	$71.00	Sign In Sign In
M0202L	400,000 units/ml	100,000 units	$281.00	$281.00	Sign In Sign In
M0202M	2,000,000 units/ml	100,000 units	$281.00	$281.00	Sign In Sign In

Please enter a quantity for at least one size

Now available as a Glycerol-free reagent. Learn more

Now available as a GMP-grade reagent. Learn more

Need a custom/large volume order? Contact us

Product Information

For details on NEB's quality controls for DNA ligases, visit our Ligase Quality page.

Bar graph of nuclease contamination in NEB lots of T4 DNA Ligase versus lots from other companies — 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:: DNA Ligases

This product can be used in the following applications:: BioBrick^® Assembly,; Cloning Ligation,; NEBridge® Golden Gate Assembly

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

M0202S

-20

	T4 DNA Ligase	M0202SVIAL	-20	1 x 0.05 ml	400,000 units/ml
	T4 DNA Ligase Reaction Buffer	B0202SVIAL	-20	1 x 1 ml	10 X

M0202T

-20

	T4 DNA Ligase	M0202TVIAL	-20	1 x 0.01 ml	2,000,000 units/ml
	T4 DNA Ligase Reaction Buffer	B0202SVIAL	-20	1 x 1 ml	10 X

M0202L

-20

	T4 DNA Ligase	M0202LVIAL	-20	1 x 0.25 ml	400,000 units/ml
	T4 DNA Ligase Reaction Buffer	B0202SVIAL	-20	1 x 1 ml	10 X

M0202M

-20

	T4 DNA Ligase	M0202MVIAL	-20	1 x 0.05 ml	2,000,000 units/ml
	T4 DNA Ligase Reaction Buffer	B0202SVIAL	-20	1 x 1 ml	10 X

Properties & Usage
Materials Required but not Supplied
- Nuclease-free Water (NEB #B1500)
Unit Definition
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

Heat Inactivation
65°C for 10 minutes
Advantages and Features
Application Features
- Cloning of restriction fragments
- Joining linkers and adapters to blunt-ended DNA
Related Products
Companion Products
- Quick Blunting™ Kit
Materials Sold Separately
- T4 DNA Ligase Reaction Buffer
Product Notes
1. ATP is an essential cofactor for the reaction. This contrasts with E. coli DNA ligase which requires NAD.
2. To dilute T4 DNA Ligase that will subsequently be stored at -20°C, 50% glycerol storage buffer (Diluent Buffer A (with rAlbumin), NEB #B8532S) should be used; to dilute for immediate use, 1X T4 DNA Ligase Reaction Buffer can be used.
3. Ligation can also be performed in any of the four restriction endonuclease NEBuffers or in T4 Polynucleotide Kinase Buffer if they are supplemented with 1 mM ATP.
4. Room Temperature Ligation
  For convenience, ligations may be done at room temperature (20-25°C). For cohesive (sticky) ends, use 1 µl of T4 DNA Ligase in a 20 µl reaction for 10 minutes. For blunt ends, use 1 µl of T4 DNA Ligase in a 20 µl reaction for 2 hours or 1 µl high concentration T4 DNA Ligase for 10 minutes. Alternatively, NEB's Quick Ligation Kit (#M2200S) [30 reactions] or (#M2200L) [150 reactions]) is uniquely formulated to ligate both blunt and cohesive (sticky) ends in 5 minutes at room temperature.
References
1. Engler, M.J. and Richardson, C.C. (1982). P.D. Boyer(Ed.), 5, 3. San Diego: Academic Press.
2. Remaut, E., Tsao, H. and Fiers, W. (1983). Gene. 22, 103-113.
3. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd Ed.). 1.53-1.73.