[Cell-biology] Re: weird problem in transfecting 293 cells in 24-well plate

Ozan Aygun via cellbiol%40net.bio.net (by metugenetics from yahoo.com)
Thu Jun 14 19:41:05 EST 2007


Hi,

we experienced that with Lipofectamine 2000 reagent,
293 cells better to be at least 90% confluent before
any transient transfection. This gives as good as
60-70 % transfection efficiency. If the cells are less
confluent than 80%, then they tend to detach
immediately after lipofectamine addition and after 48h
you may not find any living cell because of the
toxicity.

Hence make sure that your cells are confluent before
transfection. Moreover, make sure you are adding
Lipo-DNA mix from the edge of the plate, strictly not
directly over the cells and slowly -drop by drop. Be
gentle to the cells after addition of the reagent and
do not ever shake.

Hope this may be any help.

Ozan


       
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