For added convenience and accuracy in your ligation
reaction setup, try our new
DNA Ligase Master Mixes. These pre-mixed, ready-to-use
formulations include a proprietary ligation reaction enhancement agent for improved performance.
For your convenience, master mixes are available for every end type:
For sticky ends, try our Instant Sticky-end Ligase Master Mix (
NEB #M0370) For blunt or single-base overhang ends, try our Blunt/TA Ligase Master Mix (
For the full list of DNA Ligases and Ligase Master Mixes offered by NEB, as well as suggested applications, visit
DNA Ligase Selection Chart.
For information about properties of DNA and RNA Ligases, visit the Properties of DNA and RNA Ligases Chart.
For additional help, visit our
Ligase Troubleshooting Guide
For details on NEB's quality controls for DNA ligases, visit our
is a guide for selecting appropriate products and viewing protocols for steps in the cloning workflow. To help select the right ligase, choose "Ligation" on the tool to start.
Blunt/TA Ligase Master Mix improves yields for ends that typically react slowly
Yields of final ligation product for all reaction conditions using high concentration T4 DNA Ligase ( NEB #M0202), the Quick Ligation Kit ( NEB #M2200) and Blunt/TA Master Mix ( NEB #M0367). Nick, cohesive end and 3´ single-base overhang substrates were incubated for 15 minutes; the 5´ single-base overhang was incubated for 1 hour.
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-foldmolar 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 Without any additional incubation time, 2 µl were immediately withdrawn and used to transform a 50 µl aliquot of NEB 10-beta Competent E. coli ( NEB #C3019).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 wereproduced.
ElectroLigase reactions are complete in 60 minutes or less
Ligation reactions containing equal amounts (20 ng vector and 3-fold molar excess of insert) of blunt (A) or T/A (B)
vector/insert pairs were set up using ElectroLigase and incubated for the times shown. After heat inactivation of the
ligase, 2 μl of each reaction were withdrawn and directly used to transform NEB 10-beta Electrocompetent E. coli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) was plated onto selective plates and incubated
overnight at 37°C. Colonies were counted, adjusted for plating dilution, and graphed.
Electroligase® is a registered trademark of New England Biolabs.
Quick Ligation™ is a trademark of New England Biolabs
SilverXpress™is a trademark of Life Technologies, Inc.