I was doing a standard sticky-end cloning using electrocompetent cells and for some reason all the cloned inserts were larger that the gel purified DNA fragment (used methylene blue to stain the DNA in the gel and white light to visualize-so no DNA damage occurred). Looks like recombination occurred in the cells even though they were RecA1, however, there are other recombination pathways.
So I went looking for chemical competent cells to buy, which are what I normally would use, and ran across the ZYMO Research Mix & Go cells. The name says it all, just add 5 uL of ligation mixture to the 100 ul of thawed cells (with optional incubation on ice for 2-5 min.) and then add to pre-warmed plates with antibiotic! Made my life easier, and they are highly competent, really only needed to plate 10-20 uL of the 105 uL of cells/ligation mix in my case. And more importantly, all the clones had the correct size insert this time.
DH5Alpha- E.coli ... we use to make them competent by using CaCl2 .
I make my own chemically competent DH5 alpha E. coli. They will keep at -80 for about a year (we usually run out before then anyway).
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