AAT Bioquest

What should I do if I get very few, if any, transformants when transforming large plasmids ((>10 kb) or BACS?

Posted January 20, 2023


Although cells that are chemically efficient are simpler and easier to use, the one drawback with these cells is their inefficiency at taking up larger plasmids. Using electro-competent cells may be the best way to work around this drawback when you want to transform large plasmids. This involves applying an electro-magnetic field to the cell/DNA mixture to improve membrane permeability. When you need to transform large plasmids, this technique works better than using heat-shock to cause the cells to take up the DNA. You will need access to an electroporator and the appropriate cuvettes to implement this technique.

Additional resources

Properties affecting transfer and expression of degradative plasmids for the purpose of bioremediation

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Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Broad Dynamic Range*