Optimization is essential for any RNAi screen. The siGENOME Optimization Plates for Human or Mouse are ideal for selection of the best transfection conditions and controls for screens in human or 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 cyclophilin B in Human (NM_000942), Mouse (NM_011149), and Rat (NM_022536)
- Four non-targeting siRNAs and a non-targeting pool designed to have at least 4 mismatches to known human, mouse, and rat genes
|Row||Pre-plated control||Control Catalog No.|
|A||siGENOME Non-targeting siRNA #2||D-001210-02|
|B||siGENOME Non-targeting siRNA #3||D-001210-03|
|C||siGENOME Non-targeting siRNA #4||D-001210-04|
|D||siGENOME Non-targeting siRNA #5||D-001210-05|
|E||siGENOME Non-targeting pool #2||D-001206-14|
|F||siGENOME Cyclophilin B control pool (H, M, R) Control siRNA||D-001136-01|
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.
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
- B. D. Parsons et al., A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One. 4(12), e8471 (29 December 2009).
- M. Jiang et al., Tales from an academic RNAi screening facility; FAQs. Brief Funct Genomics. 10(4), 227-237 (Epub 28 April 2011, July 2011). [doi: 10.1093/bfgp/elr016]
- 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).
|Storage Conditions||4 C|
|Stability at Recommended Storage Conditions||At least 12 months|