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Transfection Efficacy


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#1 dcch

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Posted 17 November 2009 - 03:59 PM

Hello all. I would like to perform transfection optimization using oligofectamine. Invitrogen only have established protocol against HeLa and HUVEC cell line. When designing the method for this optimization, I discovered that most of the kit / paper used fluorophore labelled non targetting siRNA to perform the optimization.

I wonder if I can use the positive control (validated Lamin A/C siRNA) to carry out transfection optimization?Then assuming the Lamin knockdown as my transfection efficacy, I would produce the transfection efficacy data using reverse-transcription pcr (or WB) ?

Thank you for any suggestion!

#2 Dr Teeth

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Posted 18 November 2009 - 11:49 AM

Hello all. I would like to perform transfection optimization using oligofectamine. Invitrogen only have established protocol against HeLa and HUVEC cell line. When designing the method for this optimization, I discovered that most of the kit / paper used fluorophore labelled non targetting siRNA to perform the optimization.

I wonder if I can use the positive control (validated Lamin A/C siRNA) to carry out transfection optimization?Then assuming the Lamin knockdown as my transfection efficacy, I would produce the transfection efficacy data using reverse-transcription pcr (or WB) ?

Thank you for any suggestion!


Better to use a labeled oligo. Your second option tells you more about the knockdown efficiency than the transfection efficiency.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 dcch

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Posted 18 November 2009 - 06:59 PM

Thx for the suggestion! :rolleyes:




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