Medium for electroporating HEK 293 T-cells - (Feb/01/2008 )

Our lab recently received a Bio-Rad Gene Pulser II apparatus from a now defunct lab and it is my responsiblity to draw up a protocol for basic HEK293 T cells. I am wondering if anyone has a succesful buffer that they used during the electroporation procedure itself.

Another question, is their a significant difference between chilling the cells and cuvette or just leaving them at room temperature.

Thanks
Justin

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We used to electroporate BHK cells in PBS and immediately after pulsing them, add warm DMEM with 10% serum.

We used to chill the cuvettes and the cells were also stored on ice.

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Thanks
There doesn't seem to be a difference between chilling the cells and cuvettes and leaving them at room temperature. I thought that leaving the cells at room temperature would make the membranes more fluid, and therefore more apt to recover from the pulse. I will probably try both variables and see which one works best.

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