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Silencer® siRNA
Screening Control Panel
Effective Controls for RNAi Screening Experiments
• Convenient panel of 7 negative
control siRNAs
• Simplifies finding the best negative controls for a particular screen
• Functionally proven to have minimal effects on cell proliferation and
viability
• Validated for use in human, mouse, and rat cells
• Also includes a positive control siRNA with a clear cytotoxic phenotype
for assessing siRNA delivery efficiency
Researchers performing siRNA screens often
employ hundreds or even thousands of siRNAs per experiment. As
with experiments using individual siRNAs, effective negative
and positive control siRNAs are essential for determining transfection
efficiency and to control for the general effects of siRNA delivery
in these screens. Negative control siRNAs are also critical for
setting the “hit” threshold that determines whether
an siRNA is considered as having a positive, negative, or neutral
effect in a particular assay. Because of their importance, and
due in part to the fact that some negative control siRNAs may
have unintended effects in a particular assay, most researchers
carefully test their negative control siRNAs before choosing
ones for inclusion in their screen.
Ambion now offers the Silencer® siRNA
Screening Control Panel, a set of seven unique negative control
siRNAs that enable RNAi researchers to identify the best negative
control(s) for their cell system. These siRNAs lack similarity
to nucleotide sequences in all known human, mouse, and rat genes.
They have been tested in cell-based screens and proven to have
no significant effect on cell proliferation, cell viability,
and cell morphology, highlighting their usefulness as negative
controls. An example of the effect of transfecting these controls
on cell number is provided in Figure 1A.
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Figure
1.Effects on Relative Cell Number after
Transfections with Silencer® Screening Control
siRNAs. UMR106 (A) and HeLa (B) cells
were reverse transfected with 30 nM siRNA [Silencer Negative
Control siRNAs (A) and Silencer Kif11
siRNA (B)] in triplicate using siPORT™ NeoFX™ Transfection
Agent. Three days after transfection, the cells were
harvested, and the relative number of cells in each sample
was measured in a fluorescence plate reader using a fluorescein
diacetate substrate.
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As an added benefit, a positive control siRNA
to human, mouse, and rat Kif11 (Eg5) is included. Knockdown of
Kif11, which encodes a kinesin family motor protein, leads to
mitotic arrest (Figure 1B). Effective delivery of Kif11 siRNA
can thus be easily assessed visually; it can also be measured
by several cell-based assays.
Each of the seven negative control siRNAs,
as well as the Kif11 siRNA, in the Silencer siRNA Screening
Control Panel is provided dried at
1 nmol. Each control siRNA is also available individually at 5 nmol and can
be custom packaged to provide larger amounts.
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