Recently, I have got a problem in my electroporation. Previously, performing electroporation had been successful. But suddenly, since several months ago, all experiments failed and no clones grown after electroporation (
Try your experiment with some commercially available electrocompetent cells and using a small size plasmid as control. Also check the purity of your plasmid DNA. alternatively make electrocompetent cells and use freshly without storing. Try different protocols for making electrocompetent cells.
If previous trials were successful and trial conditions have not changed there is no reason for failure this time.
Problem solved. Thanks for your helpful suggestions, guys. After meaningful discussion with member of technical support team from Bio-Rad , I was suggested to calibrate the electroporator because the Bio-Rad machine maintenance has been discontinued since the start of the CoV19 epidemic. In addition, during the basic system test, I found that no matter what electric solutions I used the machine detected the sample resistance to be >1100ohms, suggestive of a disconnection of the electric circle in the machine. This was fixed by finding that the disconnection is due to incomplete closure of the Shock Pod chamber lid. A variety of tricky problems accompany our PhD students' daily life! Finally, I would show my great appreciation to the support from Timothy Ho and the CTS team from Bio-Rad FAS. Thanks. Jinlong Yu Kafeel Ahmad Ifeanyichukwu Okeke
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