Protocol Online logo
Top : Forum Archives: : Molecular Biology

Electroporation-Growth on Neg. Controls - (Aug/01/2002 )

Hi-I've been having difficulty with growth on my negative controls (no DNA added) from electroporation and I've been told that this sometime happens due to the high numbers of cells plated.  One of my plates has essentially a lawn of cells though--is this common?  I'm using E. coli that has the linear DNA degradation genes suppressed and is being selected on kanamycin or chloramphenicol BTW.

-Laura

-lvalent1-

If I understood you are electroporating bacteria and not cells (right?).
I have never electroporate bacteria, but use a classic method of transformation by thermic shock  (20 min on ice, 2min at 37C, and 30 min on ice). It works well and I never had more than 5 colonies growing.

-cecile-