TNT Digestion-Ligation Buffer Formulation

Our invention allow for multiple DNA fragments, inserted in a plasmid DNA, to be combined in a one-pot reaction without previous manipulation of the input DNAs (as by polymerase chain reaction linearization or gel-purification of digested fragments) with an efficient exchange of inserts toward the desired format of the recombinant DNA. Our invention saves labor-time, require no specific set of plasmids with recombination/overlap sequences, no post-reaction manipulation/clean-up is necessary, and can be 10-fold or more efficient than the most efficient method available in the market. As a consequence, subcloning/homemade competent cells (efficiency of ~10e6cfu/ug of DNA) can be used to retrieve positive clones, reducing even more the costs.

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