Blunt/TA Ligase Master Mix is a ready-to-use solution of T4 DNA Ligase, proprietary ligation enhancer, and optimized reaction buffer. This master mix is specifically formulated to improve ligation and transformation of both blunt-end and single-base overhang substrates. The master mix format simplifies reaction set-up, ensures an optimized ratio of enzyme and buffer components, and yields robust, rapid ligation of all types of DNA ends using a short incubation time at room temperature. No thawing is necessary as it remains liquid during storage at -20°C.* Ligations for subcloning can be carried out in small volumes with low DNA concentrations, allowing users to conserve precious DNA samples and directly transform many strains of chemically competent E. coli without dilution. Learn more about Joining Difficult to Ligase dsDNA Fragments and 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.* Freezers vary in their actual internal temperature. Our testing demonstrates that the master mix is liquid at -20°C. Freeze-thaw testing at -70°C has confirmed that the performance is unchanged after 20 freeze/thaw cycles.Ligation reactions with single-base overhangvector and insert were set-up using the Blunt/TA Ligase Master Mix andincubated for different times at 25°C (A) or at different temperaturesfor 15 minutes (B). Two microliters of each reaction were used totransform a 50 μl aliquot of NEB 10-beta Competent E. coli (NEB#C3019). Transformants resulting from triplicate plating 50 μl of a 1:5dilution of the outgrowth were counted and graphed. The resultsindicate that the Blunt/TA Ligase Master Mix works well at 25°C, and iscomplete in 15 minutes.
Product Source
Purified from an E. coli strain containing a recombinant gene encoding T4 DNA Ligase.
This product is related to the following categories:
DNA Ligases Products
This product can be used in the following applications:
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