Transfection Efficacy - FAM-labelled NT or Lamin? (Nov/17/2009 )
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!
-dcch-
dcch on Nov 17 2009, 06:59 PM said:
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!
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.
-Dr Teeth-
Thx for the suggestion! 
-dcch-