Optimization is essential for any RNAi screen. The ON-TARGETplus Optimization Plates are ideal for selection of the best transfection conditions and controls for screens in mouse cells using RTF siRNA Libraries. The plate layout enables the testing of cell density and transfection reagent amount on the same plate while also identifying the best non-targeting siRNA control for your assay.
- Three ready-to-use replicate 96-well plates; 6.25 pmol siRNA per well (50 nM final concentration)
- Provided in clear or clear-bottomed white or black-walled plates to support assays involving fluorescent or luminescent detection
- Positive control pool targets mouse cyclophilin B (Accession NM_011149)
- Four non-targeting siRNAs and a non-targeting pool with ON-TARGETplus modifications to minimize off-target effects
|Row||Pre-plated control||Control Catalog No.|
|A||ON-TARGETplus Non-targeting siRNA #1||D-001810-01|
|B||ON-TARGETplus Non-targeting siRNA #2||D-001810-02|
|C||ON-TARGETplus Non-targeting siRNA #3||D-001810-03|
|D||ON-TARGETplus Non-targeting siRNA #4||D-001810-04|
|E||ON-TARGETplus Non-targeting pool||D-001810-10|
|F||ON-TARGETplus Mouse Cyclophilin B control pool||D-001820-20|
DharmaFECT transfection reagents are highly recommended for use with RTF libraries and should be purchased separately. Refer to the DharmaFECT Cell Type Guide to find the appropriate formulation for your cell type.
Our siRNA knockdown guarantee
siGENOME and ON-TARGETplus siRNA reagents (SMARTpool and three of four individual siRNAs) are guaranteed to silence target gene expression by at least 75% at the mRNA level when demonstrated to have been used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 to 48 hours after transfection using 100 nM siRNA).
Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25 nM working concentration.
Optimization Plate Layout
The pre-plated controls in the RTF Optimization Plates permit assessment of multiple cell densities and transfection reagent amounts to optimize your RNAi screening conditions. You can additionally evaluate multiple non-targeting control siRNAs to determine the best negative control for your assay.
Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries
Publications using RTF Libraries
- T. Sorkina, M. Miranda, K.R. Dionne, B.R. Hoover, N.R. Zahniser, A. Sorkin, RNA interference screen reveals an essential role of Nedd4-2 in dopamine transporter ubiquitination and endocytosis. J Neurosci. 26(31), 8195-205 (2006).
- P. Monteiro, D. Gilot, E. Le Ferrec, C. Rauch, D. Lagadic-Gossmann, O. Fardel, Dioxin-mediated up-regulation of aryl hydrocarbon receptor target genes is dependent on the calcium/calmodulin/CaMKIalpha pathway. Mol Pharmacol. 73(3), 769-77 (Epub 18 December 2007, March 2008).
- A. A. Kolokoltsov, D. Deniger, E. H. Fleming, N.J. Roberts Jr, J. M. Karpilow, R. A. Davey, Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus. J Virol. 81(14), 7786-800 (Epub 9 May 2007, July 2007).
- K. M. Hussain, K. L. Leong, M. M. Ng, J. J. Chu, The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71. J Biol Chem. 286(1), 309-321 (Epub 18 October 2010, 7 January 2011).
General Screening References
- B. D. Parsons, A. Schindler, D. H. Evans, E. Foley, A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One. 4(12), e8471 (2009).
- M. Jiang, R. Instrell, B. Saunders, H. Berven, M. Howell, Tales from an academic RNAi screening facility; FAQs. Brief Funct. Genomics. 10(4), 227-237 (2011). [doi: 10.1093/bfgp/elr016]
The most broadly applicable DharmaFECT formulation for optimal siRNA or microRNA transfection into a wide range of cell types for successful RNAi experiments