ElectroLigase™ combines T4 DNA ligase and an optimized, ready-to-use 2X reaction buffer containing a proprietary ligation enhancer and no PEG. This combination is specifically formulated to promote robust ligation of all types of DNA ends (blunt, sticky, TA). It is directly compatible, without desalting or purification, with electrocompetent cells used for transformation by electroporation. No thawing of the buffer is required as it maintains a liquid state during storage at -20°C*, thereby simplifying reaction set-up. By providing an optimized ratio of enzyme and buffer components, users are able to rapidly ligate all types of DNA ends applying a short incubation time at room temperature. Ligations for subcloning can be carried out in small volumes with low concentrations, allowing users to conserve precious DNA samples. These reactions can be pipetted directly, without purification or dilution, to transform many strains of electrocompetent E. coli**.* Freezers vary in their actual internal temperatures. Our testing demonstrates that the enzyme and buffer remain liquid at -20°C. **ElectroLigase is also compatible with chemically competent strains of E. coli. Performance is generally around 50% efficiency, when compared to the Blunt/TA Ligase Master Mix (NEB #M0367 ).Ligation reactions containing equal amounts (20 ng vectorand 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 timesshown. After heat inactivation, 2 μl of each reaction were withdrawn anddirectly used to transform NEB 10-beta Electrocompetent E. coli (NEB #C3020). 50 μl aliquots of the outgrowth(diluted, in some cases) were plated onto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 incubatedovernight at 37°C. Colonies were counted, adjusted for plating dilution, andgraphed.
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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