siGENOME RTF Optimization Plates - Human/Mouse

Optimize conditions and select controls for your RTF screen


Three 96-well plates with pre-plated controls in Reverse Transfection Format (RTF) for optimization of conditions for your RNAi screen with siGENOME RTF Libraries.
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.

Highlights

  • 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
See Figure 1 (Supporting data tab) for the Optimization Plate layout.
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

Experimental considerations

DharmaFECT Cell Culture Reagent (DCCR) is recommended for use with RTF libraries to dilute transfection reagents prior to use. DCCR may be purchased separately or accompanying your RTF Optimization Plates

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.

  
HazardousNo
Shipping ConditionAmbient
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition4 C
Optimization Plate Layout

Optimization Plate Layout

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.


RTF Screening Workflow

Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries

RTF Screening Workflow


References

  1. B. D. Parsons et al., A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One4(12), e8471 (29 December 2009).
  2. 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]

Citations

  1. 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).
  2. 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).
  3. 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).
  4. 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).