Prepare Competent Cells with Speed and Efficiency!

The RapidTransit™ Transformation Kit (R2653) provides an economical and streamlined method to prepare (Figure 1) chemically competent Escherichia coli cells (Figure 2) at efficiencies suitable for standard cloning. RapidTransit reagents eliminate complex buffer preparation and lengthy incubation periods, while reducing the number of procedural steps.

Features and Benefits Fast, economical preparation of competent cells Overcome costs and strain limitations of commercially available cells Suitable for a variety of E. coli strains Eliminate complex buffer preparation Efficiencies suitable for cloning

This kit has been used successfully to prepare competent cells from a wide variety of standard and specialized E. coli strains (Figure 2), including popular K12 cloning strains (GC5™, GC10™, JM109, HB101, DH5a™, and NovaBlue™) and B strains (the BL21 family) used for recombinant protein expression.

The RapidTransit Transformation Kit offers two simple procedures (Figure 1) 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.

Kit Components
2x RapidTransit Transformation Buffer, 2.5 ml
Recovery Medium, 50 ml

Figure 1. Protocols for preparation of competent E. coli using the RapidTransit Transformation Kit

Figure 2. Comparison of Transformation Efficiency Using Different E. coli Strains. 


Competent cells were prepared using the Direct Addition method, for the strains shown above. Cells were transformed with one nanogram pUC18. Error bars indicate standard deviation calculated for the mean of triplicate reactions. A similar pattern of efficiency is observed for cells prepared by Single-Centrifugation method, though approximately 10X higher transformation efficiencies were obtained (data not shown). Transformation efficiency is expressed as colony forming units per µg plasmid.

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