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Stability of transient shRNA transfection

shRNA

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#1 schaapje_86

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Posted 17 July 2014 - 01:45 AM

When I would transiently transfect (with lipofectamine RNAimax)  my cells (Hepa1c1c7 mouse hepatoma cell line) with shRNA, how long will this transfection last and is it stable enough to last one cell passage? The reason why I am asking this is because I would like to seed my cells in a 96-well plate and I don’t want to end up transfecting each and every single well. If it isn’t possible to split cells after transfection for re-seeding purposes, is it possible to transfect cells in suspension?

 

Thanks a lot for your help!



#2 GNANA

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Posted 19 July 2014 - 11:51 AM

i think there is a protocol to transfect cells in suspension, but i would still go with transfecting them in a big plate, and split them in the destination plate around 24-36 hrs of transfection, i would keep the end point of the expt to be at 72 hrs or maximun 96 hrs of transfection.

 

good luck..


I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....





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