Human ON-TARGETplus RTF - Cell Cycle Regulation

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


A ready-to-use reverse transfection format RNAi screening library targeting human cell cycle regulation genes. Just resuspend pre-dispensed siRNA, and add cells. Optimization plates are available.

The Human ON-TARGETplus Cell Cycle Regulation library includes SMARTpool siRNA reagents targeting genes whose functions are crucial for controlling cell division. Of particular importance to the progression of the cell cycle are the cyclin-dependent kinases, or CDKs.

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
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

For a diagram of the RTF Library Plate layout, see Figure 1 on the Supporting Data tab.

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
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. General Screening References
  2. B. D. Parsonset al., A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay.PLoS One.4(12), e8471 (29 December 2009).
  3. M. Jianget 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. Publications using RTF Libraries
  2. T. Sorkinaet al., RNA interference screen reveals an essential role of Nedd4-2 in dopamine transporter ubiquitination and endocytosis.J Neurosci.26(31), 8195-8205 (2 August 2006).
  3. P. Monteiroet al.,O. Dioxin-mediated up-regulation of aryl hydrocarbon receptor target genes is dependent on the calcium/calmodulin/CaMKIalpha pathway.Mol Pharmacol.73(3), 769-777 (Epub 18 December 2007, March 2008).
  4. A. A. Kolokoltsovet al., 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),7786-7800 (Epub 9 May 2007, July 2007).
  5. K. M. Hussain KMet al., 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).