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ccdB competent cells - (Feb/06/2007 )


we use ccdB cells in our lab but the 10 Rxns are used up very fast. And they are very expensive.
Now think about to make these cells by ourself. Does anybody have experience with making these cells?



I dont have experience making these cells, but our technician makes them as any other cells and they are fine. But they tend to degrade over a period of time even after storing them in -80C.


I agree with Scolix that there is nothing special about making these cells competent. But at the very least you can make those cells last 3x as long by doing 15 ul transformations, which work just as well as 50 ul transformations, and are 3x cheaper, assuming you buy the cells. The only problem with this is that refreezing cells loses factors of 3x or so transformation efficiency. Remember to add very small amounts of your ligation product. I'd recommend 5% or less of the cell volume.