Human ON-TARGETplus RTF - DNA Damage Response

Ideal for labs who want to carry out siRNA screens but lack high-throughput capabilities.


Ready-to-use arrayed siRNA library in single-use plate sets. Just rehydrate, and add cells for RNAi screening of human DNA damage response genes. Optimization plates available.

The Human ON-TARGETplus RTF DNA Damage Response siRNA library targets genes involved in the DNA damage and repair pathways. This includes damage surveillance, damage recognition and signaling, signal effector genes, repair genes, and genes involved in dissociation and resolution of DNA repair complexes.

RTF siRNA libraries are provided as multiple single-use plate sets – just rehydrate and add cells. This unique pre-plated format reduces hands-on time for faster screening results.

ON-TARGETplus modifications combine sense strand inactivation with a novel seed-region modification. This patented siRNA approach is the best strategy to prevent off-target effects caused by both the sense and antisense strands while maintaining high silencing potency.

Highlights

  • Six ready-to-use 96-well plate sets provided for two triplicate screens
  • Pre-plated, validated RNAi controls included
  • No aliquoting necessary, just resuspend and add cells
  • siRNA reagents provided in clear plates at 6.25pmol per well (50nM final screening concentration)
  • Black or white clear-bottom plates available to support assays involving fluorescent or luminescent detection
For a diagram of the RTF Library Plate layout, see Figure 1 on the Supporting Data tab.
Well Pre-plated control Control Catalog No.
A1 ON-TARGETplus Non-targeting siRNA #1 D-001810-01
B1 ON-TARGETplus Non-targeting siRNA #2 D-001810-02
C1 ON-TARGETplus Non-targeting siRNA #3 D-001810-03
D1 ON-TARGETplus Non-targeting siRNA #4 D-001810-04
E1 ON-TARGETplus Non-targeting pool D-001810-10
F1 ON-TARGETplus Human Cyclophilin B control pool D-001820-10

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 Guideto find the appropriate formulation for your cell type.

Gene Targets

For a complete list of target genes in this siRNA Library please email Technical Support or call 1-800-235-9880. International customers please call 303-604-9499 or your local Sales Representative.

  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Storage Condition4 C

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 used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 to 48 hours after transfection using 100nM siRNA).

Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25nM working concentration.

Plate layout of ON-TARGETplus RTF siRNA Libraries

Plate layout of ON-TARGETplus RTF siRNA Libraries

Plate layout of ON-TARGETplus RTF siRNA Libraries

Validated control siRNAs and pools are pre-dispensed into column 1 of each RTF Library plate, providing a consistent baseline for screening and assay efficiency.


Reverse Transfection Format is highly effective across cell lines

Reverse Transfection Format is highly effective across cell lines

Reverse Transfection Format is highly effective across cell lines

Eight cell lines were transfected with control siRNAs targeting Cyclophilin B using RTF plates. Good cellular viability and effective target silencing were accomplished under optimized conditions. Gene knockdown is normalized to GAPDH expression. Cell plating densities are indicated in parentheses. Gene expression was determined 48 hours posttransfection by branched DNA assay (Panomics Quantigene® Reagent System) (blue bars). Cell viability was determined by alamarBlue® metabolic assay (gold circles).


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

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

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


References

  1. Publications using RTF Libraries
  2. 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).
  3. 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, March2008).
  4. 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).
  5. 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 January2011).
  6. General Screening References
  7. 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).
  8. 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]