WIXLIB

PRODUCT BROCHURE Real-Time PCR SystemsStepOne and StepOnePlusReal-Time PCR Systems Remarkably Simple Systems. Simply Remarkable Results.

Step Up To High Performance Real-Time PCRRemarkably Simple SystemsApplied Biosystems, a leader in technology solutions for life science research, introducesits latest innovations in real-time PCR—the StepOne (48-well) and StepOnePlus (96-well)Real-Time PCR Systems. These remarkably simple real-time PCR systems are specificallydesigned with a user-friendly, yet powerful interface for both new and experiencedreal-time PCR researchers.

Features At A GlanceStepOne SystemThroughput/Wells FAM /SYBR Green dyes VIC /JOE dyes ROX dyeStepOnePlus System4896 NED/TAMRA dyes VeriFlex Block Figure 1. Software HomepageSystem FeaturesThe StepOne and StepOnePlus Systems bring advancedreal-time PCR technology to a new level of accessibility—evenfor first-time users. Both systems measure amplification asit occurs, cycle by cycle, allowing for precise and quantitativemeasurements during the exponential phase of PCR. Beginningat the homepage (Figure 1), you can navigate seamlesslythrough all aspects of the real-time PCR process includingsample and reaction set-up, thermal cycling, and fluorescentdetection. Focused application software analyzes and interpretsexperimental results. Depending on the experimental design,the system can even help you select and order real-time PCRreagents online by means of convenient links in the software(optional). The system provides two-fold discrimination with99.7% confidence, along with 9 logs of dynamic range. Highlysensitive, these systems detect 10 copies of RNaseP in a 30-μLreaction. Additionally, for laboratories with the StepOne Systemthat require greater throughput, an upgrade kit is available.*StepOne UpgradeFor those customers whose lab needs are changing, AppliedBiosystems offers the StepOne System upgrade. Just sendin your existing system and you’ll receive a StepOnePlusinstrument until your upgraded system arrives in your lab.Easy-to-Use Experimental Design WizardBoth systems provide an experimental Design Wizard toassist you in setting up your first real-time PCR experiment.Just click on the Design Wizard and start answering questionsabout the real-time PCR experiment you wish to perform. TheWizard walks you through quantitation methods, detectionchemistry, ramp speeds, and nucleic acid template types.From standards to plate layout, the Design Wizard guides theentire process, including reaction mix and standard dilutioncalculations. You’re now ready to explore the simply remarkablefeatures of these systems.*StepOnePlus System Upgrade Kit coming soonSystem HighlightsFor both novice and experienced researchers, the StepOne orStepOnePlus Systems have what you need. Cost-effective, 3-color/48-well (StepOne) or 4-color/96-well(StepOnePlus) Systems deliver precise, quantitative real-timePCR results Long-life LED-based optical system records fluorescencefrom FAM /SYBR Green I, VIC /JOE , and ROX dyes forgene expression analysis, pathogen quantitation, SNPgenotyping, and presence/absence assays; Note: TheStepOnePlus System also accommodates the TAMRA dye Both systems perform standard and fast PCR reactions inless than 40 minutes Ultra-compact footprint fits any laboratory setting LCD touchscreen and USB drive provide configurationflexibility and enable PC free operation Remote monitoring and email notification for convenienceand time-savings The StepOnePlus System features VeriFlex Blocktechnology, which combines six independentlycontrollable peltier blocks for enhanced PCRfunctionality and precise temperature control The StepOne System is upgradeable to the StepOnePlusSystem to meet your changing research needs

Simply Remarkable InterfaceFlexibilityBoth the StepOne and StepOne Plus Systems adapt toalmost any workflow with flexible instrument control and datamanagement. From the touchscreen control panel, you canquick-start your experimental run without PC connectivity. Youcan also create a new protocol, view the history of your lastrun, or see protocol details. When the run is complete, datacan be downloaded easily onto a USB flash stick or savedto a PC. Either system can be connected directly to a LocalArea Network (LAN), and you can monitor the progress of theexperiment, send new instructions to the system, downloaddata, and edit the instrument profile. The software for the twosystems also contains a convenient email feature that notifiesyou when your experiment is complete and ready for analysis.The StepOne and StepOnePlus Systems can be installed inmultiple distinct configurations, providing unmatched flexibilityand convenience. The unique standalone (PC free) configurationprovides an ultra-compact footprint that will fit into any laboratory. A direct connection to a LAN enables remote monitoringof experimental progress and downloading of the completedrun file to the PC at your desk.System Configurations (Figure 4)1. PC controlled2. PC free3. Networked4. PC controlled connected to LAN5. PC controlled with networked instrumentFigure 2. Browse Experiment Page On TouchscreenFigure 3. Run Monitor Screen On TouchscreenFigure 4. Five Distinct Configurations1. PC controlled2. PC free3. NetworkedLAN

SoftwareThe StepOne and StepOnePlus Systems software containsunique features not available in other real-time PCR instruments. For example, data can be analyzed from multipleperspectives in the Multiple Plots view (See Figure 5). Thesoftware constructs four-plot, side-by-side views of all dataaspects including the amplification plot, standard curve,multi-component data plots, and raw data. It also displays thisdata next to the plate layout for easier analysis. Another novelfeature is the software’s ability to automatically identify wellsthat might compromise the success of an experiment. Duringdata analysis, the software generates a quality-control reporttable that flags wells based on criteria such as amplificationin a negative-control well, the absence of a signal in a well, ora high CT standard deviation in a replicate group. This featurereduces analysis time and allows even researchers new toreal-time PCR to have confidence in their results. Furthermore,you can customize this feature by disabling or modifyingquality flag settings to suit your experimental needs. Tools for easy identification of sample wells whenviewing amplification curves or SNP genotyping plots Email notifications to alert you when a run has startedor ended Easy cut-and-paste functionality Export easily to PowerPoint , Excel , or directly as a .jpeg fileFigure 5. StepOne System Multiple Plots ViewSoftware Highlights Experimental Design Wizards to help you design and setup experiments Pipetting protocols and recipes to set upexperiments quickly Advanced set-up for expert users who require flexibilityfor more complex applications, such as multiplexing Quick-start set-up so you can begin a run immediatelyand enter plate information later Real-time monitoring of amplification growth curves enablesyou to view run progress (can be viewed from a remote PC) Auto-baseline and auto-threshold for simplified data analysis Multiple Plots view for simultaneous data assessment fromfour perspectives Automated SNP genotype-calling with intuitive graphicaloutput and quality-value assignment Troubleshooting flags to help you diagnose and solveproblematic experimentsFigure 6. StepOne System Reaction Set-up ViewFor each target in the reaction plate, the StepOne System software lets youreview and edit calculated volumes for preparing standards and PCR reactions.4. PC controlled connected to LANUSB—10/100 Adapter5. PC controlled with network instrumentLAN

Simply Remarkable ApplicationsInstrument ChemistriesBoth StepOne Systems are equipped with fluorophoredetection chemistries that include FAM and VIC dye-labeledTaqMan MGB probe-based assays, VIC and TAMRA dyelabeled probe-based assays and SYBR Green I dye chemistry.[Note: the TAMRA dye is available only with the StepOnePlusSystem.] TaqMan probe-based assays provide outstandingspecificity and sensitivity, and SYBR Green I dye chemistry is aneconomical alternative for target identification, initial screeningassays, or assays that require only a few reactions.Software AnalysisThe software for the StepOne Systems supports a varietyof analysis methods, including: Melt curve analysis Genotyping (including real-time amplification)ApplicationsThe StepOne Systems support any real-time PCRapplication. Predesigned or custom assays exist for thefollowing applications: SNP Genotyping Translocation Analysis Gene Expression Profiling Gene Detection MicroRNA Expression Viral Load AnalysisFor information about existing Gene Expression, MicroRNA andTranslocation Analysis Assays, please visit www.allgenes.comAbsolute Quantitation- Standard curveSNP Genotyping Assay information can be found atwww.allsnps.comRelative Quantitation- Relative standard curve- Comparative CT (ΔΔCT)SpeedThe StepOne and StepOnePlus Systems perform bothstandard and fast thermal cycling on the same block with nomodification. Standard thermal cycling requires less than twoPresence/absence (plus/minus) assays with an internalpositive controlFigure 8. Genotyping AnalysisFigure 7. Broad Dynamic RangeAmplification PlotAllele Discrimination Plot (SNP Assay: SNP Assay 1)104.13.63.12.6ΔRnAllele 212.11.61.10.60.10.10.024681012141618202224 262830323436380.20.40.60.81.01.21.41.61.82.0Allele 140CycleHomozygous 1/1Amplification plot shows log of change in normalized reporter fluorescenceplotted vs. PCR cycle number. This plot from the StepOne System illustrates 9logs of linear dynamic range for a TaqMan Assay of cDNA containing the 18Starget sequence in tenfold serial dilutions.Heterozygous 1/2Homozygous 2/2UndetermindedStepOne and StepOnePlus software automatically determines genotypes andgenerates an intuitive graphic representation of results in a cluster plot reportthat helps you view data across populations or samples. Results are from humanCYP2C19*2 TaqMan SNP Genotyping Assay (using the StepOnePlus System).

hours, and fast thermal cycling significantly reduces the runtimes of quantitative real-time PCR applications by deliveringresults in 40 minutes. Fast cycling is ideal for maximizing thenumber of runs on an instrument in any given work day.VeriFlex BlocksThe 96-well StepOnePlus System features VeriFlex Block technology, whichbrings six independently controllablepeltier blocks together for precisetemperature control and enhancedPCR functionality. VeriFlex Blocksdeliver flexibility for those who haveprobes and primers that are optimizedat different annealing temperatures.VeriFlex Block (96-well)ResultsBecause the StepOne and StepOnePlus Systems are factorycalibrated for optical and thermal accuracy, simply remarkablereal-time PCR results are available right out of the box. Bothsystems can discriminate between two populations of 5,000and 10,000 template copies of a TaqMan assay with 99.7%confidence. Both systems also demonstrate a linear dynamicrange of 9 log units or more, as shown by the amplificationplot (See Figure 7).Figure 10. Multiplex AssayFigure 9. Melt Curve AnalysisMultiplex AssaysMelt Curve103.42.411.9ΔRnDerivative Reporter (-Rn’)2.91.40.1.9.4- .10.0165.070.075.080.0Tm: 79.285.090.0Temperature ( C)In this experiment, standard PCR was run to generate amplicons for 18S,ß-actin and GAPDH. Aliquots of each amplicon were combined into onereaction tube and a melt curve analysis was performed.95.00246810 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42CycleMultiplex TaqMan Assays on the StepOnePlus System showingamplification of cDNA (96 samples) using probes labeled with VIC and FAM reporters for 18S and TGF-ß target sequences respectively.