Products for RNAi

From design to detection,Bio-Rad supports

your RNAi research.

RNAi Solutions

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siLentMer siRNA (27 nt)

Dicer

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Target recognitionand cleavage

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2 Products for RNAi

RNAi –– Powerful, Specific Gene SilencingRNA interference (RNAi) is one of the most powerful and widely used techniques available for the specific inhibition of gene expression. Using RNAi, researchers can specifically and potently modulate the expression of target genes. By analyzing resulting changes in gene expression at both the protein and RNA level, researchers are rapidly advancing the understanding of gene function.

Performing RNAi experiments involves several steps, and at each step it is important to eliminate variables, use appropriate controls, and work with reagents and instrumentation that provide sensitive, reliable results. Bio-Rad offers a broad selection of reagents, instrumentation, and expertise to support you through virtually every step of your RNAi workflow.

The RNAi PathwayRNAi is a naturally occurring mechanism for genesilencing found in all eukaryotic cells and is believedto have biological roles in gene regulation anddefense from viral attack. The RNAi pathway isactivated by double-stranded RNAs (dsRNAs). Long dsRNAs are cleaved by Dicer endonuclease, an RNase III family member, to form 21–23 ntduplexes known as short interfering RNAs (siRNAs).After cleavage, siRNA duplexes are incorporated intoan RNA-induced silencing complex (RISC). Dicer, inaddition to cleaving dsRNAs, may facilitate this step.After incorporation into RISC, the siRNA duplex is unwound and the guide strand retained in theenzyme complex. The RISC complex, via strandhomology, pairs with an mRNA target which is then cleaved by RISC.

Activation of the RNAi Pathway In the laboratory, the RNAi pathway can be activatedby expression of short hairpin siRNA (shRNA) usingplasmid or viral vectors, by transfection of in vitrotranscribed dsRNAs, or by transfection of chemicallysynthesized siRNAs. Transfection of siRNAs isperhaps the most commonly used approach, and it is favored because such siRNA substrates are simpleto generate, fast acting, and typically highly effective.

The earliest reports of RNAi activation in mammaliancells demonstrated that chemically synthesized 21–23 nt siRNAs activate the RNAi pathway at theRISC assembly step (Elbashir et al. 2001). Recentreports, however, have shown that 27 nt siRNAs,which enter the RNAi pathway at the Dicer cleavagestep, can be up to 100-fold more potent than 21-mersiRNAs containing the same sequence (Kim et al.2005, Rose et al. 2005). This effect is likely due toDicer having a role in RISC assembly and subsequentpreferential strand loading. These 27-mer siRNAs arecalled Dicer-substrate siRNAs.

Dicer-substrate siRNAs can be designed toconsistently promote specific cleavage of a singlesiRNA product by Dicer and direct retention of the strand complementary to the target mRNA. These siRNAs mediate effective knockdown at low concentrations, thereby reducing the potential for the off-target effects typically associated with high siRNA concentrations.

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27-mer Dicer-Substrate siRNA — More Potent Gene SilencingPresynthesized, ready-to-order siLentMer siRNA duplexes eliminate the time, effort, and expenseassociated with designing, synthesizing, and testingmultiple specific siRNAs.

Designed against a variety of commonly studied targets,these potent 27-mer Dicer-substrate siRNAs enter the RNAi pathway earlier than traditional 21–23-mersiRNAs (see opposite page). As a result, siLentMer siRNAduplexes can mediate effective silencing at extremely lowconcentrations.

siLentMer validated and predesigned siRNA duplexes offerthe following advantages:

� ≥85% knockdown with ≥5 nM siRNA (validated)� Reduced potential for off-target effects � HPLC-purified for consistent and reliable results

siLentMer Validated siRNA Duplexes

All siLentMer validated siRNA duplexes are functionallytested to ensure a minimum silencing efficiency of ≥85% using ≥5 nM siRNA. These highly effective siRNAduplexes are ideal for silencing a target gene or for use as positive and negative controls. Up to two validateddesigns are available for each target gene to allowconfirmation of specific silencing; a fluorescently labelednonsilencing control siRNA is also available.

siLentMer Predesigned siRNA Duplexes

siLentMer predesigned siRNA duplexes are ideal when a specific siLentMer validated siRNA is not available.Although these siRNAs are not validated, they aredesigned using the same advanced algorithm used for the validated siRNA duplexes. For each gene target, up to four designs are available.

For a complete list of available targets, visit us on the Web at www.bio-rad.com/RNAi/ or contact your localBio-Rad representative.

Effective siRNAs and Controls

Potent, well-designed siRNAs are crucial to the specific silencing of a targetgene. Bio-Rad, partnering with IDT, employs the novel Dicer-substrate siRNAtechnology to provide a collection of validated and predesigned siRNAduplexes for specific, effective silencing using as little as 5 nM siRNA.

Products for RNAi 3

siLentMer siRNA duplexesmediate effective knockdown with low concentrations of siRNA.HeLa cells were transfected with 10 nM or 100 pM siLentMer anti-GAPDH siRNA in a 12-well plateusing 0.6 µl siLentFect™ lipid reagentper well. At 48 hr posttransfection,total RNA was extracted and RT-qPCR performed using the iScript™

cDNA synthesis kit and the iCycleriQ® real-time detection system. In both cases, the level of GAPDHtranscript ( � ) was reduced by >95%relative to a nonsilencing control ( � ).

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Lipid-Based Transfection siLentFect Lipid Reagent for RNAi

siLentFect lipid reagent is a high-efficiency, low-toxicitycationic lipid specifically designed to deliver siRNA to a broad variety of cultured mammalian cells for RNAiapplications. The exceptional affinity of siLentFect forsiRNA and its efficient delivery of complexed siRNA conferrobust targeted gene silencing using low lipid volumes andlow concentrations of siRNA. With siLentFect lipid reagent,you can achieve:

� Exceptional transfection of a broad range of cell lineswith minimal cytotoxicity

� 90–99% silencing of high- and low-abundance targetswith as little as 100 pM siRNA

� Efficient transfection with low volumes of lipid

� Transfection of adherent and suspension cells

� Codelivery of siRNA and double-stranded DNA vectorsfor optimization of RNAi experiments

� Effective tissue array-based reverse transfectionexperiments

A Choice of Delivery Strategies

A successful RNAi experiment requires that siRNA or expression vectors be delivered to asmany cells as possible with minimal impact on cell health. For many cell types, lipid-basedtransfection reagents meet these requirements, but for cells resistant to lipid-basedtransfection, other methods, such as electroporation or biolistic delivery, may be used.

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Efficient transfection with low volumes of lipid and low concentrations of siRNA. CHO cells stablyexpressing the lacZ gene were grown in 24-well platesand transfected with 10 nM of an siRNA targetinglacZ. Cells were lysed and assayed for β-galactosidaseactivity 24 hr posttransfection (left). A significantreduction in expression of lacZ was observed evenwith low volumes of siLentFect (right).

4 Products for RNAi

10 nM100 pM

siLentFect enables effective silencing with low concentrations of siRNA.HeLa cells were transfected with siLentFect reagent and 10 nM or 100 pM27-mer siRNAs targeted against three different reference genes. Cells werelysed 24 hr posttransfection and total RNA was isolated using the Aurum™

total RNA mini kit. Expression levels of the three genes were measured by RT-qPCR and compared to the levels in cells transfected with a nonsilencingcontrol siRNA.

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Examples of Successfully Transfected Cell Lines

A549 C166-GFP Caco-2CHO-K1 COS-7 HEK 293

HeLaHUVECJurkat NIH 3T3LNCaPMCF-7

Primary fibroblastPrimary keratinocyteSCC12B23T3-Swiss albino

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TransFectin™ Lipid Reagent — Transfection of shRNA Plasmidsand General-Purpose Transfection

This powerful cationic lipid reagent delivers the highestpossible efficiencies to a broad range of cell lines. In RNAistudies, TransFectin is appropriate for transfection of shorthairpin RNA (shRNA) expression plasmids and cassettes.For the most commonly used cell lines, TransFectinsurpasses the performance of other high-efficiencycationic lipid reagents with reduced cytotoxic effects.TransFectin lipid reagent offers:

� Successful delivery to a broad range of cells

� Exceptional transfection efficiencies at 40–90% culture confluence

� Efficient transfection in the presence of serum-containing media

� Effective transfection with less lipid for a reduced cost per transfection

ElectroporationGene Pulser Xcell™ Electroporation System

With numerous published references, the Gene Pulser Xcell electroporationsystem is proven for siRNA and DNAdelivery into primary, suspension, and difficult-to-transfect cells.

Key features include:

� Tunable delivery for difficult-to-transfect cells

� Square waveforms for gentle delivery to sensitive cells

� User-friendly interface, and choice of preprogrammedprotocols or manual operation

� Complete report of conditions applied to cells

� Patented* PulseTrac™ circuitry for reproducibility andsample protection

Biolistic Particle DeliveryHelios® Gene Gun and PDS-1000/He™ Systems

Biolistic technology, or particlebombardment, is a physical method ofdelivering nucleic acid into cells that canbe applied to the widest range of targets,including eukaryotic cell cultures, tissues,

and organs. In RNAi experiments, this technology isparticularly useful for cell types, such as plant cells, that cannot be transfected using lipid-based orelectroporation methods.

The Helios gene gun and the PDS-1000/He systems useadvanced biolistic technology to transfect cells either in vitro or in situ. These systems:

� Facilitate both transient and stable expression� Require small amounts of nucleic acid� Are capable of codelivery of multiple plasmids

* US patents 4,750,100 and 4,910,140.

Products for RNAi 5

Silencing from a transfected expression vector. HeLa cells weretransfected with pSilencer anti-GAPDH plasmid (Ambion, Inc.) usingTransFectin lipid reagent. At 48 hr posttransfection, total RNA was extractedand GAPDH mRNA levels were quantitated by RT-qPCR (left). Levels ofGAPDH transcript in transfected ( � ) cells were reduced by ~90%compared to untransfected cells ( � ) (right).

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Harvest cells

Quantitate protein yield (RC DC™ protein assay kits)

Clean/concentrate extract (ReadyPrep 2-D cleanup kit)

Resolubilize extract (2-D compatible buffer)

Reduce and alkylate protein for 2-D(ReadyPrep reduction-alkylation kit)

2-D separation and analysis

RNA Isolation Aurum Total RNA Kits

Careful RNA preparation is important to ensure high-quality RNA that is free of contaminants, such as PCR inhibitorsor genomic DNA, that can affectdownstream results. Aurum total RNA kits

isolate very pure RNA that meets the demanding needs ofRT-qPCR and microarray-based analysis. Efficient andeasy to use, these kits are available in spin- or vacuum-compatible formats to streamline sample preparation. These kits provide:

� A safe aqueous isolation procedure� High yields of RNA from many sample types� Ready-to-use RNA (DNA- and phenol-free)� Quick protocol using silica membrane binding� RNase-free reagents, plasticware, and DNase I

Protein Sample Preparation ReadyPrep™ Kits

The success of protein-based analysis of gene silencingdepends on sample purity. Many protein extraction andsolubilization techniques introduce contaminants, such as salts, detergents, and ionic compounds, that caninterfere with protein separation. Eliminating suchcontaminants ensures the success of western blotting or two-dimensional (2-D) electrophoresis.

Bio-Rad offers a suite of protein sample preparationproducts. For the purpose of protein extraction fromcultured cells after transfection of siRNA, a recommendedworkflow is shown below. For more information aboutprotein sample preparation products to use for RNAi, visit www.bio-rad.com/RNAi/. For more informationabout other protein sample preparation products, visit www.expressionproteomics.com

The Importance of Sample Quality

The results of RNAi are most commonly assessed at the mRNA level through reverse-transcriptionquantitative PCR (RT-qPCR) or microarray-based approaches, or at the protein level through westernblotting, 2-D electrophoresis, protein arrays, or functional analyses. Regardless of the detectionmethod, the purity of the mRNA or protein sample is critical to success. Bio-Rad offers a selection ofeffective sample preparation and assessment products to help you achieve the best results possible.

Analysis of high-quality RNA isolated with the Aurum total RNA fatty andfibrous tissue kit. Left, electropherogram of high-quality mouse brain totalRNA analyzed on the Experion™ automated electrophoresis system using the Experion RNA StdSens analysis kit. Right, denaturing agarose gelelectrophoresis of RNA samples (4 µg) from mouse brain and skeletal muscle.

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Impact of siRNA-targeted RNAsample quality on RT-qPCR. Experionelectropherograms of intact or partiallydegraded total RNA from transfectionswith scrambled control siRNA and anti-GAPDH siRNA are shown. Panels A andB show RT-qPCR traces obtained usingthe combination of scrambled control( � ) and anti-GAPDH ( � ) RNA samplesindicated in the electropherograms(positioned above and beside thetraces). Also shown are the averagemeasured CT values for each sampleand the overall level of gene silencing,which is indicated by the ΔCT value.Assessment of RNA quality is criticalbecause poor or variable sample qualitymay skew results, as shown by the shiftin CT of 2.5 in the degraded controlsample, potentially leading todisregarding effective siRNAs or keeping siRNAs that exert only weakmRNA suppression.

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Sample Assessment Experion Automated Electrophoresis System

Following sample preparation, the quality of experimentaland control samples should be assessed prior toperforming downstream analysis. The integrity of an RNAsample, for example, is a crucial parameter when verifyingthe specificity and extent of siRNA-induced gene silencing(Perez-Novo et al. 2005, Strong and Rubio 2005).

Though total RNA and protein samples may be evaluatedby traditional gel-based electrophoretic methods, thesemethods are time-consuming and may require significantamounts of precious samples. The Experion automatedelectrophoresis system utilizes microfluidic technology to combine the numerous steps associated withelectrophoresis, from separation through imaging andanalysis. The Experion system combines the utility

and qualitative benefits of gel-based analysis with thecapabilities of quantitative spectroscopic analysis to provide:

� Accurate qualitative analysis, sizing, and quantitation of purified RNA and protein samples

� Analysis of 10–12 samples in just 30 minutes

� Minimal sample and reagent volume requirements

� Automatic calculations, including some statisticalanalysis

� Intuitive navigation and powerful data comparison tools

For more information, request bulletin 3140 or go to www.bio-rad.com/experion/

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Products for RNAi 7

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Real-Time RT-qPCR RT-qPCR is the fastest, most reproducible, and mostsensitive technique for specific mRNA detection andquantitation currently available. In RT-qPCR, cDNAs aregenerated from mRNA templates by reverse transcriptionand amplified by PCR. Amplification is observed in realtime, enabling accurate quantitation of PCR products.

Bio-Rad provides a comprehensive selection of real-timePCR instrumentation and reagents for single and multiplextarget analysis.

Real-Time PCR Instruments

The iQ™5 and MyiQ™ real-time detectionsystems are modular upgrades that add real-time PCR capabilities to the iCycler®

thermal cycler. The iQ5 is a five-targetmultiplex real-time instrument offered with

a complete software package for experimental design and analysis. The MyiQ is a powerful single-color detectionsystem ideal for detection of common green fluorescentdyes, such as FAM and SYBR Green I.

Bio-Rad also offers instrumentation for single- andmulticolor fluorescence detection that is based onlong-lived LED illumination platforms. These systems take advantage of sequential excitation and detection to minimize cross talk and maximize sensitivity.

Reverse Transcription and Real-Time PCR Reagents, iScript cDNA Synthesis Kits

RT-qPCR can be accomplished in one ortwo steps. In two-step reactions, reversetranscription is carried out separately from

PCR, allowing several real-time PCR assays to be performedon a single cDNA sample. In one-step reactions, the reversetranscription and qPCR processes are combined into asingle reaction tube to offer ease of use and minimizeexperimental variation (Wong and Medrano 2005).

� iScript and iScript™ Select cDNA synthesis kitsrepresent a breakthrough in first-strand cDNA synthesis— streamlined protocols that require only 45 minutes

� iScript one-step quantitative RT-PCR kits are versatileenough for use with any fluorescent detection chemistry

Gene Expression Analysis

Gene silencing can be monitored at the mRNA level using a variety ofdifferent techniques. Real-time RT-qPCR has become the preferredmethod for validating gene-specific knockdown, and microarrays are used to evaluate gene expression changes on a global scale.

The iScript cDNA synthesis kit performs across a broad range of RNA concentration. HeLa cellRNA was reverse-transcribed with the iScript cDNAsynthesis kit and then the β-actin target was amplifiedusing iQ SYBR Green supermix. The iScript cDNAsynthesis kit delivers at least 6 orders of dynamicrange while maintaining optimum sensitivity with limitedamounts of input RNA.

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Supermixes for PCR and Real-Time PCR

Bio-Rad’s supermixes are premixed blends of buffer,dNTPs, and polymerase. Specially formulated to generateunrivaled results in real-time PCR, they are qualified for use on a variety of real-time thermal cycler platforms.Supermixes are available in five formulations: iQ™ supermix,iQ™ SYBR® Green supermix, iTaq™ SYBR® Green supermixwith ROX, iTaq™ supermix with ROX, and for multiplexqPCR, iQ multiplex powermix.

PCR Enzymes and dNTP Mix

Bio-Rad's iTaq DNA polymerase is an antibody-mediated,hot-start polymerase that ensures high specificity andsensitivity for both conventional and real-time PCRapplications. The companion dNTP mix is ready for use and is qualified for real-time PCR.

Microarray AnalysisMicroarray Chip Arrayer

For screening applications that require the monitoring of more than just a fewtargets, microarray-based analysis isrecommended. Bio-Rad’s powerful

BioOdyssey™ Calligrapher™ miniarrayer platform mediates both gene expression andprotein screening for RNAi applications,and enables fast and accurate protein,cDNA, and RNA printing.

Analysis of changes in protein expression cancomplement and confirm data generated fromanalysis of mRNA knockdown. Western blots, 2-D electrophoresis, and protein arrays can all beused to monitor changes in protein expressionresulting from RNAi-mediated knockdown of aspecific gene target.

Western Blotting Western blot detection is a convenient method for the assessment of gene silencing. In western analysis,proteins are separated by polyacrylamide gelelectrophoresis (PAGE) and then transferred to amembrane for detection by specific antibodies. A leadingprovider of protein gel electrophoresis instrumentation and reagents, Bio-Rad offers a trusted line of products for electrophoresis, blotting, detection, and analysis:

� Efficient, versatile vertical electrophoresis and blotting cells

� Easy-to-use precast polyacrylamide gels

� Convenient premixed buffers and precut blotting membranes

� A wide range of recombinant and natural protein standards

� Sensitive chemiluminescent and colorimetricimmunodetection kits

Protein Expression Analysis

Western analysis of gene silencing.HeLa cells were transfected with 5 nM of a 27-mer control or anti-β-actin siRNA. Cells were harvested 48 hrposttransfection and lysed in Laemmlisample buffer. Cell proteins (20, 8, and 2 µg) were separated by SDS-PAGE in a 12% Ready Gel®

precast gel, transferred to nitrocellulose,and detected colorimetrically usingantibodies specific to β-tubulin (upper panel) and β-actin (lower panel).Note knockdown of β-actin expressionrelative to control.

Products for RNAi 9

Control siRNA20 8 2 µg

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2-D ElectrophoresisWith the ability to resolve complex mixtures of proteinssimultaneously in a single gel, 2-D electrophoresis is one of the most powerful protein separation techniquesavailable. In 2-D electrophoresis, proteins are firstseparated by isoelectric focusing (IEF) and then by SDS-PAGE. This approach allows not only the analysis of the discrete reduction of protein expression as a result of targeted silencing, but also the visualization ofbroad changes in protein expression. This enables theidentification of up- and downregulated proteins that maybe interrelated.

Bio-Rad offers a complete selection of products forprotein sample preparation, 2-D analysis, and imaging for expression proteomics. To learn more about Bio-Rad’s products for 2-D electrophoresis, visitwww.expressionproteomics.com

Suspension ArraysBio-Plex™ Suspension Array System

The Bio-Plex system is a bead-based fluorescent multiplexarray system that provides another powerful tool forassessment of protein expression following gene silencing.The Bio-Plex system uses up to 100 color-coded beadsets, each of which can be conjugated with a differentreactant, to permit the simultaneous analysis of up to 100 different proteins in a single microplate well. By multiplexing with the Bio-Plex system, you candramatically increase the amount of useful informationfrom rare or volume-limited samples.

The Bio-Plex suspension array system offers a detectionplatform, sophisticated software package, comprehensiveand premixed phosphoprotein and cytokine assays, andcustomized x-Plex™ assays to simplify differential proteinexpression analysis from cell lysates.

� Simultaneously quantitate up to 100 protein targets in culture media, sera, and other matrices

� Instantly customize your assays by mixing bead sets

� Automatically analyze up to 96 samples in under 35 minutes

Detection (cont.)

10 Products for RNAi

2-D analysis of β-actin silencing. HeLa cells were transfected with a nonspecific control or 5 nM anti-β-actin siRNA. Cells were lysed 48 hrposttransfection, and protein was extracted and then separated by 2-Delectrophoresis. Image analysis showed a 3–5-fold reduction in β-actinprotein (top panels) and a 2-fold increase in expression of other proteins(lower panels, labeled 1 and 2) in cells transfected with anti-β-actin siRNA.

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Bio-Rad is dedicated to providing tools and technologies that enable the application and interpretation of results generated from the use of RNAi. We provide a range oftechnologies for RNAi, from design through detection and analysis of both mRNA and protein.

As a customer-focused, technology-based company, we continue to develop enablingtechnologies for RNAi and related applications. Our goal is to provide you with thetools and technologies you need, and we invite your comments and ideas. To contactus, or to learn more about Bio-Rad’s RNAi-enabling technologies, visit us on the Webat www.bio-rad.com/RNAi/ or contact your local Bio-Rad office.

ReferencesElbashir SM et al., Duplexes of 21-nucleotide RNAs mediate RNAinterference in cultured mammalian cells, Nature 411, 494–498 (2001)

Kim D-H et al., Synthetic dsRNA Dicer substrates enhance RNAipotency and efficacy, Nat Biotechnol 23, 222–226 (2005)

Perez-Novo CA et al., Impact of RNA quality on reference geneexpression stability, Biotechniques 39, 52, 54, 56 (2005)

Rose SD et al., Functional polarity is introduced by Dicer processing of short substrate RNAs, Nucleic Acids Res 33, 4140–4156 (2005)

Strong W and Rubio T, Using the Experion automated electrophoresissystem to assess RNA quality and quantity in siRNA-induced genesilencing experiments, Bio-Rad bulletin 5315 (2005)

Wong ML and Medrano JF, Real-time PCR for mRNA quantitation,Biotechniques 39, 75–85 (2005)

Notice regarding Bio-Rad thermal cyclers and real-time systems.Purchase of this instrument conveys a limited non-transferableimmunity from suit for the purchaser’s own internal research anddevelopment and for use in applied fields other than Human In VitroDiagnostics under one or more of U.S. Patents Nos. 5,656,493,5,333,675, 5,475,610 (claims 1, 44, 158, 160–163 and 167 only), and6,703,236 (claims 1–7 only), or corresponding claims in their non-U.S.counterparts, owned by Applera Corporation. No right is conveyedexpressly, by implication or by estoppel under any other patent claim,such as claims to apparatus, reagents, kits, or methods such as 5'nuclease methods. Further information on purchasing licenses may beobtained by contacting the Director of Licensing, Applied Biosystems,850 Lincoln Centre Drive, Foster City, California 94404, USA.

Bio-Rad’s real-time thermal cyclers are licensed real-time thermalcyclers under Applera’s United States Patent No. 6,814,934 B1 foruse in research and for all other fields except the fields of humandiagnostics and veterinary diagnostics.

pSilencer is a trademark of Ambion, Inc. SYBR is a trademark ofMolecular Probes, Inc. Bio-Rad Laboratories, Inc. is licensed byMolecular Probes, Inc. to sell reagents containing SYBR Green I foruse in real-time PCR, for research purposes only.

The IDT logo is a trademark of Integrated DNATechnologies, Inc. These products are sold underlicense of U.S. Patent No. 6,506,559 (Carnegie)

and pending and issued patents of Alnylam Pharmaceuticals, Inc., and pending patents of Integrated DNA Technologies, Inc. for researchuse only. Not for use as a therapeutic or diagnostic in humans,animals, or plants. For custom siRNA synthesis, contact IDT.

LabChip and the LabChip logo are trademarks of CaliperLife Sciences, Inc. Bio-Rad Laboratories, Inc. is licensed byCaliper Life Sciences, Inc. to sell products using the

LabChip technology for research use only. These products arelicensed under US Patent Nos. 5,863,753, 5,658,751, 5,436,134,and 5,582,977, and pending patent applications, and related foreignpatents, for internal research and development use only in detecting,quantitating, and sizing macromolecules, in combination withmicrofluidics, where internal research and development use expresslyexcludes the use of this product for providing medical, diagnostic, orany other testing, analysis, or screening services, or providing clinicalinformation or clinical analysis, in any event in return for compensationby an unrelated party.

The Bio-Plex suspension array system includes fluorescently labeledmicrospheres and instrumentation licensed to Bio-Rad Laboratories,Inc. by the Luminex Corporation.

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Life ScienceGroup

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Bio-Rad Laboratories, Inc.

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Ordering InformationCatalog # Description

Design

siLentMer Dicer-Substrate siRNA DuplexesVaries siLentMer Validated Dicer-Substrate siRNA Duplexes,

2 nmol, designed with proven criteria and functionallytested for ≥85% silencing

Varies siLentMer Predesigned Dicer-Substrate siRNA Duplexes,2 nmol, designed with proven criteria

siLentMer Delivery Optimization Kit174-9950 siLentMer Delivery Optimization Kit, includes 1.0 nmol

fluorescently labeled siLentMer nonsilencing siRNA, 0.2 ml siLentFect lipid reagent, 1.0 ml siLentMer siRNAresuspension buffer

siLentMer Starter Kits174-9960 siLentMer Starter Kit for Human GAPDH, includes

0.5 nmol siLentMer validated human GAPDH siRNA(positive control), 0.5 nmol siLentMer nonsilencing siRNA(negative control), 0.2 ml siLentFect lipid reagent, 1.0 mlsiLentMer siRNA resuspension buffer

174-9961 siLentMer Starter Kit for GFP, includes 0.5 nmolsiLentMer validated GFP siRNA (positive control), 0.5 nmol siLentMer nonsilencing siRNA (negative control),0.2 ml siLentFect lipid reagent, 1.0 ml siLentMer siRNAresuspension buffer

siLentMer Total Control Kits*174-9970 siLentMer Total Control Kit for Human GAPDH174-9971 siLentMer Total Control Kit for Human HPRT174-9972 siLentMer Total Control Kit for Human Lamin A/C174-9973 siLentMer Total Control Kit for Human Cyclophilin174-9974 siLentMer Total Control Kit for Human β-Actin174-9975 siLentMer Total Control Kit for Human β-Tubulin174-9976 siLentMer Total Control Kit for GFP174-9977 siLentMer Total Control Kit for Luciferase

* All total control kits include 1.0 nmol siLentMer validated siRNA (positive control), 1.0 nmol fluorescently labeled siLentMer nonsilencingsiRNA (control for delivery), 1.0 nmol siLentMer nonsilencing siRNA(negative control for silencing), 0.5 ml siLentFect lipid reagent, 1.0 mlsiLentMer siRNA resuspension buffer

Go to www.bio-rad.com/RNAi/ for a complete list of catalog numbers for validated and predesigned siRNAs and control kits.

Delivery170-3360 siLentFect Lipid Reagent for RNAi, 0.5 ml170-3361 siLentFect Lipid Reagent for RNAi, 1.0 ml170-3362 siLentFect Lipid Reagent for RNAi, 5 x 1.0 ml170-3350 TransFectin Lipid Reagent, 0.5 ml170-3351 TransFectin Lipid Reagent, 1.0 ml170-3352 TransFectin Lipid Reagent, 5 x 1.0 ml165-2660 Gene Pulser Xcell Total System165-2661 Gene Pulser Xcell Eukaryotic System165-2086 Gene Pulser/MicroPulser™ Cuvettes, 0.2 cm gap, 50165-2088 Gene Pulser/MicroPulser Cuvettes, 0.4 cm gap, 50165-2431 Helios Gene Gun System, 100/120 V165-2257 PDS-1000/He System

Go to www.bio-rad.com/genetransfer/ for more information about Bio-Rad’s gene transfer product offering.

Catalog # Description

Sample Purification — RNA732-6800 Aurum Total RNA 96 Kit, 2 x 96-well preps732-6820 Aurum Total RNA Mini Kit, 50 preps732-6830 Aurum Total RNA Fatty and Fibrous Tissue Kit, 50 preps732-6370 AquaPure™ RNA Isolation Kit, 100 preps

Sample Purification — Protein161-0737 Laemmli Sample Buffer, 1x, 30 ml500-0121 RC DC Protein Assay Kit I, 450 standard assays500-0122 RC DC Protein Assay Kit II, 450 standard assays163-2130 ReadyPrep 2-D Cleanup Kit, 50 preps163-2105 ReadyPrep 2-D Starter Ki163-2090 ReadyPrep Reduction-Alkylation Kit, 100 preps

Sample Assessment700-7000 Experion Automated Electrophoresis System,

100–120/220–240 V, for protein analysis700-7001 Experion Automated Electrophoresis System,

100/120 V, for RNA analysis700-7002 Experion Automated Electrophoresis System,

200/240 V, for RNA analysis700-7101 Experion Pro260 Analysis Kit for 10 Chips700-7103 Experion RNA StdSens Analysis Kit for 10 Chips700-7105 Experion RNA HighSens Analysis Kit for 10 Chips

Detection and Analysis of RNA by Real-Time qPCR170-8890 iScript cDNA Synthesis Kit, 25 x 20 µl reactions170-8892 iScript One-Step RT-PCR Kit With SYBR Green,

50 x 50 µl reactions170-8860 iQ Supermix, 100 x 50 µl reactions170-8880 iQ SYBR Green Supermix, 100 x 50 µl reactions170-8850 iTaq SYBR Green Supermix With ROX, 200 x

50 µl reactions170-8848 iQ Multiplex Powermix, 50 x 50 µl reactionsCFB-3120 MiniOpticon™ Real-Time PCR System, includes optical

housing, MJ Mini™ thermal cycler, analysis software170-9770 MyiQ Single-Color Real-Time PCR Detection System170-9780 iQ5 Multicolor Real-Time PCR Detection System

Microarrayers169-2000 BioOdyssey Calligrapher MiniArrayer, 115 V169-2100 BioOdyssey Calligrapher MiniArrayer, 230 V169-2040 Humidity Control Module (HCM), for BioOdyssey

Calligrapher miniarrayer

Detection and Analysis of Protein by Western Blotting170-3930 Mini Trans-Blot® Electrophoretic Transfer Cell,

10 x 7.5 cm gel size170-4070 Criterion™ Blotter With Plate Electrodes,

15 x 9.4 cm gel size170-3939 Trans-Blot® Cell With Plate Electrodes and Super Cooling

Coil, 16 x 20 cm gel sizeGo to www.bio-rad.com/proteinblotters/ to find out more aboutmembranes and detection kits for western blot detection.

Expression Proteomics and 2-D ElectrophoresisGo to www.expressionproteomics.com for more information on Bio-Rad’s complete line of 2-D analysis products.

Multiplex Suspension Array SystemGo to www.bio-rad.com/bio-plex/x-plex/ for a complete listing ofavailable Bio-Plex assays.

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