For added convenience and accuracy in your ligation reaction setup, try our 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: Instant Sticky-end Ligase Master Mix (NEB #M0370
) for sticky ends, and Blunt/TA Ligase Master Mix (NEB #M0367
) for blunt or single-base overhang ends.
Learn more about NEB's quality controls for DNA ligases.
For help with selecting or using a DNA ligase, visit our supporting materials:
NEBcloner, an interactive tool for selecting appropriate products and viewing protocols for steps in the cloning workflow
DNA Ligase Selection Chart
Substrate-based Ligase Selection Chart
Properties of DNA and RNA Ligases Chart
Ligase Troubleshooting Guide
App note: Joining Difficult to Ligate dsDNA Fragments
App note: Efficient Adaptor Ligation for the Preparation of dsDNA Libraries using Blunt/TA Ligase Master Mix
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 Without any additional incubation time, 2 µl were immediately withdrawn and used to transform a 50 µl aliquot of NEB 10-beta Competent E. coli (NEB #C3019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 After heat inactivation of the ligase, 2 µl of each reaction were withdrawn and directly used to transform NEB 10-beta Electrocompetent E. coli
). 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.