New Method of Preparing Cells for Standard Cloning

A new way to prepare custom competent cells at efficiencies suitable for standard cloning purposes has been developed.

The method entails the use of the RapidTransit Transformation Kit, which provides an economical and streamlined way to prepare chemically competent Escherichia coli cells at efficiencies suitable for standard cloning. RapidTransit reagents eliminate complex buffer preparation and lengthy incubation periods while reducing the number of procedural steps. The kit has been used successfully to prepare competent cells from a wide variety of standard and specialized E coli strains, including popular K12 cloning strains and B strains (the BL21 family) used for recombinant protein expression.

The RapidTransit Transformation Kit offers two simple procedures to prepare competent cells: the direct addition method and the single-centrifugation method. The direct addition method allows for preparation of competent cells, transformation, and recovery within a single tube or well. The direct addition method provides transformation efficiencies of > 1 X 105 colony forming units (cfu)/µg supercoiled pUC18 plasmid DNA, whereas the single-centrifugation method provides reproducible transformation efficiencies of > 1 X 106 cfu/µg supercoiled pUC18 plasmid DNA, when using strain JM109.

"The RapidTransit system is an example of Sigma's commitment to the development of innovative products for cloning and recombinant protein expression,” said Keith Joliff, manager of global strategic marketing, molecular biology, at Sigma-Aldrich Corporation (St. Louis, MO, USA; www.sigmaaldrich.com), which developed the new system. "RapidTransit complements our current line of chemically competent and electrocompetent cells while providing researchers who require specialized strains or economical formats with a convenient alternative.”




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