MDA-MB-231 cell transfection
Posted 01 June 2011 - 07:41 AM
I've tried to transfect MDA-MB-231 with lipofectamine 2000 and it didn't work. I've been using 1ul lipofectamine for 3wells (of a 24plate) because it is the amount my labmates are using with another cell lines to avoid citotoxicity.
I've seen that invitrogen has a special lipofectamine (Lipofectamine™ LTX Reagent) for this kind of cells but it seems that you need to collect cells between 18-24hours after transfection and i would need to collect them after 48hours. Does anybody transfect these cells before and can tell me with reagent/amount has used?
Have you worked with Lipofectamine™ LTX Reagent before and know how it works?
Posted 04 June 2011 - 02:56 PM
Posted 25 June 2011 - 05:31 PM
I have used Lipfectamine, lipofectamine 2000, Lipofectamine LTX, Fugene 6 and PEI for various transfections of DNA and there is always some cytotoxicity. Personally I have seen the greatest cytotoxicity with the lipofectamine variants, and the least with PEI, but results vary between researchers.
Posted 28 June 2011 - 12:43 PM
I've been searching for some electroporation protocols, and everyone I've found is for 48 well plates or 6 well plates. I'd like to try 24well plates, due some cells would die during electroporation, i'm not sure about how many cells should i seed per well...
I'm using gene pulser Xcell from biorad in 0.2cm cuvettes, with a concentration at electroporation time of 1x106cell/ml, but no idea about how many cells should I electroporate and seed for each well. any idea?