The inoue method for preparation and transformation of competent e. Coli: "ultra-competent" cells.

DMSO Inoue transformation buffer (please see Step 1) Chilled to 0°C before use. Plasmid DNA (recombinant plasmid) Construct using one of the methods described in Directional Cloning into Plasmid Vectors, Attaching Adaptors to Protruding Termini, Blunt-ended Cloning into Plasmid Vectors, Dephosphorylation of Plasmid DNA, Addition of Synthetic Linkers to Blunt-ended DNAand Ligating Plasmid and Target DNAs in Low-melting-temperature Agarose.