We recommend the following workflow.
- dsRNA synthesis from DRSC reagent collection templates (PCR amplification, in vitro transcription, normalization)
- incubation of cells with RNAi and cell-based assay
We provide protocols for 6-well and 384-well plates, with additional tips on quantities for 96-well plates (see 384-well plate protocol). Experiments may be done in other size plates, just scale up or down, and test the assay (some adjustments might be needed).
Please note that a number of factors will influence RNAi knockdown, including
- reagent design
- reagent quality/purity
- reagent amount
- cell type
- cell density
- cell passage number (tip: don't let cells overgrow or passage them for a long time prior to the experiment--they might stop responding to RNAi reagents)
Feel free to contact us with questions about cell-based RNAi in small or large-scale studies.