I am doing an electroporation of Plasmid DNA (~1kbp) and electrocompetent E. coli DH5a. How many uL of cells and DNA should I use. I have 1uL DNA and 25uL cells per NEB protocol but I have found varying answers.
1 ul DNA / 20-25 ul cells is the standard for almost every electroporation reaction.
As Michael Swyers wrote, just use 1 uL DNA if all you need is transformed bacteria (and no quantification of efficiency etc). Depending on the competency of the cells and the DNA used, you (most likely) need to spread dilutions of your transformation to get well-separated cfus.
1 ul DNA and 25 ul cells is ideal. But you need to plate in different dilutions to get a single colony spread
Thanks to everyone for the answers! @Micheal, @Alexander, and @Sumedha thank you very much for confirming for me. I have used the aforementioned proportions and plated with dilutions with no success. I made my own electrocompetent cells so the error could have been in that portion of my procedure. I will run it again!
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