Hello Sarah,

I have had similar experiences with transformation by electroporation. The problems you are experiencing could definitely be explained by incomplete/inefficient transformation. Heat-shock transformation in TOP10 chemically competent E.coli worked better for me.

I assume you have optimized the electroporator settings for your given experiment? If not, I would try varying the amount of pulses, the voltage, and the duration of the pulse(s). If your E. coli are treated and thawed right, the problem might be that the electroporation process kills off most of the bacteria resulting in the low plasmid yields.

Best of luck.

Hello Sarah,

I had a quite similar problem with electroporation of bacterial artificial chromosome in recombinant E.Coli. I tried to increase the efficiency by increasing the washing by cold water as well as trying to keep everything as cold as possible( washing water, electporation cuvettes and the bacteria)

Best of luck

Hey Sarah, just had quick look online and it seems that plasmid has a low copy number, so that would explain the low yields. Try growing it up in TB broth instead of LB (See Wood et al 2017). Also try growing it at different temperatures 25, 30, 37 that can help with the colony formation at times. When purifying the DNA use a lower volume of elution buffer so your sample is more concentrated and spin it down twice into the same eppendorf to enhance your yield. If you happen to have the qiagen maxi prep kit it could be worthwhile to try that too. Hope that helps and best of luck!