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HomeFAQsI used an ElectroLigase® reaction to transform electrocompetent cells and I experienced arcing of my sample. What happened?
FAQ: I used an ElectroLigase® reaction to transform electrocompetent cells and I experienced arcing of my sample. What happened?
ElectroLigase reactions have been formulated to be directly compatible with electrocompetent cells. Testing on many samples does not reveal a problem using typical electroporation conditions. Arcing during electroporation can be caused by low resistance or high conductivity in the sample. The typical factors affecting conductivity during electroporation are sample temperature and salt concentration. The competent cells should be kept cold on ice and care should be taken to ensure the cuvette used does not warm up prior to being subjected to the electric field. This means wear gloves and only wipe the cuvette with a Kimwipe if necessary, taking care to not warm the sample. DNA used in ligations with ElectroLigase should be clean (e.g. column purified) and suspended in aqueous solution containing low ionic strength (10 mM Tris or less is ok). Lastly, air bubbles trapped in the sample may prevent sample contact with both electrodes of a cuvette. A few simple taps of the cuvette on a bench, after loading the DNA/comp cell mixture, can help dislodge trapped air.