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DNA Ligase Master Mixes

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Blunt/TA Ligase Master Mix

The Blunt/TA Ligase Master Mix (NEB #M0367) 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. In this application note, we demonstrate that this mix with proprietary enhancers delivers superior performance when working with blunt or single base-pair fragments.

Blunt/TA Ligase Master Mix Datacard

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.

Blunt/TA Ligase Master Mix Outperforms the Competition

Duplicate ligation reactions of blunt or T/A vector/insert pairs were set upaccording 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 (NEB #M0202), proprietary ligation enhancer and optimized reaction buffer, the Instant Sticky-end Ligase Master Mix (NEB #M0370) 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 Datacard

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.

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