2 replies to this topic

[yer_maw]

Hi,

Im looking for a protocol for electroporation of HEK293 cells and cant seem to find a particularly good one.  

What do you suspend the cells in and at what density? How much DNA do you add? How do you keep the cells nice after electroporating?

When i transform JM109 e.coli i have to pellet and wash with ice cold distilled water 4 times then resuspend in 2 volumes of water before electroporating them to remove salts. I dont see anything like this for HEK?


cheers

Edited by yer_maw, 02 April 2009 - 08:27 AM.

[genehunter]

293 cells will go burtsed in water as they dont have the cell wall struture bacteria have.   your machine will not work well on mammalian cells anyway.

293 cells can be readily transfected with CaPO4, lipid or polymer-based transfection agents and no need to use electroporation.

[Penguin]

A quick google search found this straightforward protocol

 transfection_by_electrop.pdf   56.54K   15 downloads

P