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how to compensate the cell death during transfection by electroporation


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#1 sonum

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Posted 25 November 2010 - 09:39 AM

hi
i am trying to find out a good cell cycle profile after transfection with my gene of interest by using electroporation method.but unfortunately,after transfection there is lots of cell death in control cells also(null transfection),So am getting sharp peaks in sub g1 population while cell cycle analysis in control cell.so please help me to get rid of these dead cells.so i can get a good cell cycle profile with no or very less sub g1 population.


regards
sonum

#2 Rsm

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Posted 25 November 2010 - 11:52 PM

You will always have dead cells after electroporation. That is due to the nature of electric shocks.
I would recommend using lipofection, or just culture your cells for 48h after transfection (with some washes to remove dead cells).

rsm
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#3 Denis Baev

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Posted 19 January 2011 - 08:46 AM

hi
i am trying to find out a good cell cycle profile after transfection with my gene of interest by using electroporation method.but unfortunately,after transfection there is lots of cell death in control cells also(null transfection),So am getting sharp peaks in sub g1 population while cell cycle analysis in control cell.so please help me to get rid of these dead cells.so i can get a good cell cycle profile with no or very less sub g1 population.


regards
sonum

Gate-out that cells and generate the cell cycle histogram only for cells within the live cell gate.




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