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Calculating siRNA Transfection Efficiency

siRNA transfection efficiency

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3 replies to this topic

#1 vetvinoth

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Posted 22 January 2012 - 03:38 AM

Hi all,

I am going to Transfect siRNA in primary Chicken embryo fibroblast. Please suggest me how to calculate the transfection efficiency.
I ve read about using fluorescently labelled siRNAs like Alexa flour. But presently our Flow cytometer is not working. please suggest me an alternative as i am nearing my year end of my research and i have to finish my work.

THanks....
VetVinoth

#2 pcrman

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Posted 22 January 2012 - 09:51 AM

You can also view the transfected cells under a microscope to determine how many cells are fluorescence positive.

#3 vetvinoth

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Posted 23 January 2012 - 02:00 AM

Thank you for your reply,

i also thought of counting cells but regarding calculating transfection efficiency please suggest weather i can calculate by this formula


(NO of Cells showing fluorescence in a field / total no of cells in a field ) * 100

or any other method is there...


Vinoth

#4 bob1

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Posted 23 January 2012 - 03:41 PM

That formula will give you a percentage, so it is correct for that method. If you wanted to do a lot of work determining the baselines and fluorescence levels for particular percentage transfections, you could use a fluorescent plate reader to determine the efficiency.





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