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Home > Product Overview > DNA Ligases and Ligase Master Mixes
DNA Ligases and Ligase Master Mixes
With over 37 years of experience in the development and production of enzymes for molecular biology, NEB offers the most extensive commercially available selection of high-quality, and performance-optimized DNA ligases and ligase master mixes to streamline your cloning experiments.

Benefits of DNA Ligases and Ligase Master Mixes:
  • Fast ligations
  • Robust ligation efficiency
  • Industry standard for purity
  • Most extensive selection commercially available
  • New formulations optimized for your substrates
  • Convenient master mix formats

New DNA Ligase Products

T7 DNA Ligase
Sticky-end ligation and nick sealing can be efficiently catalyzed by T7 DNA Ligase (1,2). However, unlike T4 and T3 DNA Ligases, blunt-end ligation is not efficiently catalyzed by T7 DNA Ligase, making it a good choice for applications in which blunt and sticky ends of DNA are present but only the sticky ends are to be joined.

  1. Doherty, A.J. et al. (1996) J. Biol. Chem. 271, 11083-11089.
  2. Subramanya, H.S. et al. (1996) Cell. 85, 607-615.
Sticky-end specific ligation with T7 DNA Ligase

Ligation reactions containing blunt- (ΦX174 DNA-HaeIII Digest, NEB #N3026)
and sticky-end (λ- HindIII Digest, NEB #N3012) fragments were set up with
200 ng of each substrate and 1 µl of each ligase, and incubated for 30 minutes
at 25°C in their corresponding reaction buffers. Reactions were immediately
stopped with 6X loading dye and resolved by electrophoresis on a 1% agarose
gel and stained with ethidium bromide.

T3 DNA Ligase

Sticky ends, blunt ends, and nick sealing can all be efficiently catalyzed by T3 DNA Ligase (1). As with T4 DNA Ligase, blunt-end ligation is enhanced by the addition of PEG 6000 to the reaction. T3 DNA Ligase exhibits a higher tolerance (2-fold) for NaCl in the reaction compared to T4 DNA Ligase, making the enzyme a versatile choice for in vitro molecular biology protocols requiring DNA ligase activity.

  1. Cai, L. et al. (2004) J. Biochem. 135, 397-403.

Blunt/TA Ligase Master Mix

This master mix is specifically formulated to improve ligation and transformation of both blunt-end and single-base overhang substrates. This master mix format simplifies reaction setup, ensuring robust, rapid ligation with an optimized ratio of enzyme and buffer components. Reactions can be used to directly transform many strains of chemically competent E. coli without dilution.

Blunt/TA Ligase Master Mix outperforms the competition

Duplicate ligation reactions of blunt or T/A vector/insert pairs were set up
according to the master mix vendors' suggestions. Equal amounts of ligated
DNA were used to transform NEB 10-beta Competent E. coli (NEB #C3019)
and triplicate plating was performed. Transformation results were averaged
and graphed as a percentage of the highest performing reaction, T/A ligation
using the Blunt/TA Ligase Master Mix.

Instant Sticky-end Ligase Master Mix
Complete with T4 DNA Ligase, proprietary ligation enhancer and optimized reaction buffer, the Instant Sticky-end Ligase Master Mix enables instant, incubation- free ligation. Specifically formulated to rapidly ligate sticky-end (2–4 bp) substrates and improve transformation, this mix simplifies reaction set-up and reduces the time needed for routine ligations. Reactions can be used to directly transform many strains of chemically competent E. coli without dilution.

Instant Sticky-end Ligase Master Mix offers robust ligation with no incubation step

Reactions containing 20 ng of pUC19 digested with SacI/SphI, and a 3-fold
molar excess of a 1.6 kb fragment from a different plasmid with compatible
ends were set up using the Instant Sticky-end Ligase Master Mix. Without
any additional incubation time, 2 µl were immediately withdrawn and used to
transform a 50 µl aliquot of NEB 10-beta Competent E. coli (NEB #C3019).
A 50 µl aliquot of the 1 ml outgrowth was plated onto a selective plate and
incubated overnight at 37°C. Results show that several hundred colonies were

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ElectroLigase is specifically formulated for robust ligation of all types of DNA ends
(blunt-, sticky-, T/A) and is directly compatible, without desalting or purification,
with electrocompetent cells used for transformation by electroporation.

ElectroLigase reactions are complete after 60 minutes or less, even on blunt- or T/A ends

Ligation reactions containing equal amounts (20 ng vector and 3-fold molar excess of insert) of blunt- (A), T/A (B), or sticky-end (C) vector/insert pairs were set up using ElectroLigase and incubated for the times shown. After heat inactivation, 2 μl of each reaction were withdrawn and directly used to transform NEB 10-beta Electrocompetent E. coli (NEB #C3020). 50 μl aliquots of the outgrowth (diluted, in some cases) were plated onto selective plates and incubated overnight at 37°C. Colonies were counted, adjusted for plating dilution, and graphed.

Ligase Selection Chart
Master Mix
Master Mix
ElectroLigase™ T4 DNA
E. coli DNA
Ligation of sticky ends .
Ligation of blunt
. . . . .
T/A cloning . . . .
Electroporation . . . . . . . . .
Ligation of stickyends
. . . . . . . . . .
Repair of nicks
in dsDNA
High complexity
library cloning
. . . . . . .
Ligase Detection
Reaction & Ligase
Chain Reaction
. . . . . . . . .
NGS Library Prep
. . . . . . .
Salt tolerance
(>2X that for
T4 DNA Ligase)
. . . . . . . . . .
Ligation in 15 min.
or less
. . .
Master Mix
. . . . . . . .
Thermostable . . . . . . . . .

Optimal, recommended
ligase for selected
Works well for
Will perform selected
application, but is not
Please consult the specific
NGS protocol to determine the
optimal enzyme for your needs

Blunt/TA Ligase Master Mix M0367S/L 50/250 reactions
Instant Sticky-end Ligase Master Mix M0370S/L 50/250 reactions
ElectroLigase™ M0369S 50 reactions
T4 DNA Ligase M0202S/L/M/T 20,000/10,000 units
Quick Ligation™ Kit M2200S/L 30/150 reactions
T7 DNA Ligase M0318S/L 100,000/750,000 units
T3 DNA Ligase M0317S/L 100,000/750,000 units
E. coli DNA Ligase M0205S/L 200/1,000 units
Taq DNA Ligase M0208S/L 2,000/10,000 units
9°N™ DNA Ligase M0238S/L 2,500/12,500 units
Quick Ligation Module E6056S/L 20/100 reactions

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