2 replies to this topic

[dcch]

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!

[Dr Teeth]

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!

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

[dcch]

Thx for the suggestion!