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StepOne and StepOnePlusReal-Time PCR SystemsRemarkably simple systems.Simply remarkable results.
High-performance, low-throughputreal-time PCRRemarkably simple systemsThe Applied Biosystems StepOne (48-well) and StepOnePlus (96-well) Real-Time PCRSystems represent the latest innovations in real-time PCR from a leader in technologysolutions for life science research. These remarkably simple real-time PCR systemsare specifically designed with a user-friendly yet powerful interface for both new andexperienced researchers.
System featuresThe StepOne and StepOnePlus systems bring lowthroughput real-time PCR technology to a new level ofaccessibility—even for first-time users. Both systemsmeasure amplification as it occurs, cycle by cycle, allowingfor precise and quantitative measurements during theexponential phase of PCR. Beginning at the homepage(Figure 1), you can navigate seamlessly through all aspectsof the real-time PCR process, including sample and reactionsetup, thermal cycling, and fluorescent detection. Focusedapplication software analyzes and interprets experimentalresults. Depending on the experimental design, the systemcan even help you select and order real-time PCR reagentsonline by means of convenient links in the software (optional).The system provides 2-fold discrimination with 99.7%confidence, along with 9 logs of dynamic range. These highlysensitive systems detect 10 copies of RNase P in a 30 µLreaction. Additionally, for laboratories with the StepOnesystem that require greater throughput, an upgrade kitis available.Features at a ls4896FAM/SYBR greendyes VIC/JOE dyes ROX dye NED/TAMRAdyes VeriFlex block System highlightsFor both novice and experienced researchers, theStepOne or StepOnePlus systems have what you need. Both systems offer the ability to perform high-resolutionmelt analysis (additional software package required) Cost-effective 3-color/48-well (StepOne) or 4-color/96well (StepOnePlus) systems deliver precise, quantitativereal-time PCR results Long-life, LED-based optical system recordsfluorescence from FAM /SYBR Green I, VIC /JOE ,and ROX dyes for gene expression analysis, pathogenquantitation, SNP genotyping, and presence/absenceassays. The StepOnePlus system also accommodatesthe TAMRA dye Figure 1. Software homepage.StepOnePlus Real-Time PCR System UpgradeThe Applied Biosystems StepOnePlus Real-Time PCRSystem Upgrade is designed to accommodate yourchanging real-time PCR needs. The upgrade converts aStepOne system into a StepOnePlus system by upgradingthe 48-well block to a 96-well Applied Biosystems VeriFlex Block and expanding the dye flexibility to four colors. Thecolor of the bar above the block is also updated to indicatethat the instrument is now a StepOnePlus system. Justsend in your existing StepOne system and you’ll receive aStepOnePlus loaner instrument to use until your upgradedsystem arrives in your lab.Easy-to-use experimental design wizardBoth systems provide an experimental design wizard to assistyou in setting up your first real-time PCR experiment. Justclick on the design wizard and start answering questionsabout the real-time PCR experiment you wish to perform.The design wizard walks you through quantitation methods,detection chemistry, ramp speeds, and nucleic acidtemplate types. From standards to plate layout, the designwizard guides the entire process, including reaction mix andstandard dilution calculations. You’re now ready to explorethe remarkably simple features of these systems. Both systems perform Fast PCR reactions in less than40 minutes Ultra-compact footprint fits any laboratory setting LCD touch screen and USB drive provide configurationflexibility and enable PC-free operation Remote monitoring and email notificationfor convenience and time savings The StepOnePlus system features VeriFlexBlock technology, which combines sixindependently controllable Peltier blocks for enhancedPCR functionality and precise temperature control The StepOne system is upgradable to the StepOnePlussystem to meet your changing research needs3
Simply remarkable interfaceFlexibilityBoth the StepOne and StepOnePlus systems adapt toalmost any workflow with flexible instrument control and datamanagement. From the touch screen control panel, you canquick-start your experimental run without PC connectivity.You can also create a new protocol, view the history of yourlast run, or see protocol details (Figures 2 and 3). When therun is complete, data can be downloaded easily onto a USBflash stick or saved to a PC. Either system can be connecteddirectly to a Local Area Network (LAN), and you can monitorthe progress of the experiment, send new instructions tothe system, download data, and edit the instrument profile.The software for the two systems also contains a convenientemail feature that notifies you when your experiment iscomplete and ready for analysis.The StepOne and StepOnePlus systems can be installedin multiple distinct configurations, providing unmatchedflexibility and convenience. The unique stand-alone (PC-free)configuration provides an ultra-compact footprint that willfit into any laboratory. A direct connection to a LAN enablesremote monitoring of experimental progress and downloadingthe completed run file to the PC at your desk.Figure 2. Browse experiment page on the touch screen.1. PC-controlled2. PC-freeSystem configurations (Figure 4)1. PC-controlled2. PC-free3. Networked4. PC-controlled, connected to LAN5. PC-controlled, with networked instrumentSoftwareThe Applied Biosystems StepOne and StepOnePlus Systems software contains unique features not availablein other real-time PCR instruments. For example, data canbe analyzed from multiple perspectives in the multiple plotsview (Figure 5). The software constructs four-plot, side-byside views of all data aspects, including the amplificationplot, standard curve, multicomponent data plots, and rawdata. It also displays this data next to the plate layout foreasier analysis. Another novel feature is the software’s abilityto automatically identify wells that might compromise thesuccess of an experiment. During data analysis, the softwaregenerates a quality-control report table that flags wellsbased on criteria such as amplification in a negative-controlwell, the absence of a signal in a well, or a high Ct standarddeviation in a replicate group. This feature reduces analysisFigure 3. Run monitor screen on the touch screen.3. NetworkedLANFigure 4. Five distinct configurations.
time and allows researchers new to real-time PCR to haveconfidence in their results. Furthermore, you can customizethis feature by disabling or modifying quality flag settings tosuit your experimental needs.Software highlights Optional software package offers the ability to conducthigh-resolution melt experiments View amplification plot, multicomponent plot, andQC summary within the study to easily identify andeliminate outliers Preview the effect of modified analysis settings beforepermanently applying them to results Experimental design wizards help you design and setup experiments Pipetting protocols and recipes help to set upexperiments quickly Advanced setup if you require more flexibility forcomplex applications such as multiplexing Quick-start setup so you can begin a run immediately andenter plate information later Real-time monitoring of amplification growth curvesenables you to view run progress (can be viewed from aremote PC)Figure 5. StepOne system multiple plots view. Multiple plots view for simultaneous data assessmentfrom four perspectives Remote monitoring and email notification with run fileattached for immediate access to results—even remotely Export easily to Microsoft PowerPoint or Excel software,or directly as a .jpeg filePowerful gene expression study package (Figure 6) Import an unlimited number of comparative Ct(relative quantitation) plates to one study View data by biological replicate group or technicalreplicate group Normalize to multiple endogenous controls Enter known efficiencies to adjust RQ values foreach target4. PC-controlled, connected to LANUSB—10/100 adapterFigure 6. StepOne system with the gene expression study package.The StepOne Software v2.1 includes the gene expression study package toanalyze an unlimited number of plates in one study, and displays the resultsin a publication-ready fashion.5. PC-controlled, with networked instrumentLAN
Simply remarkable applicationsInstrument chemistriesBoth StepOne systems are equipped with fluorophoredetection chemistries that include FAM and VIC dye–labeled Applied Biosystems TaqMan MGB probe-basedassays, VIC and TAMRA dye–labeled probe-based assays,and SYBR Green I dye chemistry. [note: The TAMRA dyeis available only with the StepOnePlus system.] TaqManprobe-based assays provide outstanding specificity andsensitivity, and SYBR Green I dye chemistry is an economicalalternative for target identification, initial screening assays, orassays that require only a few reactions.Software analysis (Figures 7 and 8)The software for the StepOne systems supports a variety ofanalysis methods, including: Absolute quantitation– standard curve Relative quantitation– relative standard curve– comparative Ct ( Ct) Presence/absence (plus/minus) assays with an internalpositive control Melt curve analysis Genotyping (including real-time amplification) High-resolution melt analysis (additional softwarepackage required)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 analysis Methylation Mutation scanningFor information about existing Applied Biosystems geneexpression, microRNA, and translocation analysis assays, goto thermofisher.com/allgenesSNP genotyping assay information can be found atthermofisher.com/taqmansnpSpeedThe StepOne and StepOnePlus systems perform bothstandard and Fast thermal cycling on the same block withno modification. Standard thermal cycling requires lessthan 2 hours, and Fast thermal cycling significantly reducesthe run times of quantitative real-time PCR applicationsby delivering results in 40 minutes. Fast cycling is ideal formaximizing the number of runs on an instrument in anygiven workday.Amplification plotAllele discrimination plot (SNP assay: SNP assay 1)104.13.63.12.6 RnAllele 212.11.61.10.60.10.10.02468101214 1618 202224 262830323436380.20.40.60.8CycleFigure 7. Broad dynamic range. Amplification plot shows log of changein normalized reporter fluorescence plotted vs. PCR cycle number. Thisplot from the StepOne system illustrates 9 logs of linear dynamic range foran Applied Biosystems TaqMan Assay of cDNA containing the 18S targetsequence in 10-fold serial dilutions.1.01.21.41.61.82.0Allele 140Homozygous 1/1Heterozygous 1/2Homozygous 2/2UndetermindedFigure 8. Genotyping analysis. StepOne and StepOnePlus softwareautomatically determines genotypes and generates an intuitive graphicrepresentation of results in a cluster plot report that helps you view dataacross populations or samples. Results are from human CYP2C19*2Applied Biosystems TaqMan SNP Genotyping Assay (using theStepOnePlus system).
VeriFlex BlocksThe 96-well StepOnePlus systemfeatures Applied Biosystems VeriFlex Block technology,which brings six independentlycontrollable peltier blocks togetherfor precise temperature controland enhanced PCR functionality.VeriFlex Blocks deliver flexibilityfor those who have probes andprimers that are optimized atdifferent annealing temperatures.VeriFlex Block (96-well)ResultsBecause the StepOne and StepOnePlus systems are factorycalibrated for optical and thermal accuracy, remarkablysimple real-time PCR results are available right out of the box(Figures 9 and 10). Both systems can discriminate betweentwo TaqMan Assays containing 5,000 and 10,000 copiesof template with 99.7% confidence. Both systems alsodemonstrate a linear dynamic range of 9 log units or more,as shown by the amplification plot (Figure 7).Multiplex assaysMelt curve103.42.411.9 RnDerivative Reporter (-Rn’)2.91.40.1.9.4- .10.0165.070.075.080.0Tm: 79.285.090.095.0Temperature ( C)Figure 9. Melt curve analysis. Standard PCR was run to generateamplicons for 18S, β-actin, and GAPDH. Aliquots of each amplicon werecombined into one reaction tube and a melt curve analysis was performed.0246810 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42CycleFigure 10. Multiplex assay. Multiplex TaqMan Assays on theStepOnePlus system showing amplification of cDNA (96 samples) usingprobes labeled with VIC and FAM reporters for 18S and TGF-β targetsequences, respectively.
Systems at a glance.5 cm (19depth: 48.1 in).5 cm (19.1 in)height: 51.2 cm (20.4 in)height: 51.2 cm (20.4 in)depth: 48width: 24.6cm (9.7 in)width: 24.6cm (9.7 in)StepOne systemStepOnePlus system 48 wells for lower throughput 96 wells for higher throughput 3 colors for basic applications research 4 colors for more flexibility Easily upgradable to StepOnePlus system VeriFlex Block technology for thermal cycling flexibility
Remarkable Applied Biosystems solutionsWe offer a complete range of real-time PCR reagents anddesign/analysis software to assist both experienced andnovice users of the StepOne and StepOnePlus Real-TimePCR Systems. During experimental setup, the design wizardautomatically creates a material list that links directly to ouronline store for convenient ordering of Applied Biosystems real-time PCR master mixes and consumable plasticware(optional).Service and support at every stepinstrument troubleshooting. Hands-on classes are availableat scientific training centers, in your lab, or through ourApplied Biosystems LabCoat Live virtual training.Technical supportContact our team of technical support scientists withquestions about product selection or use, assay orexperiment design, and data analysis or troubleshooting.Online support centers are available 24 hours.Online instrument managementSign in to your thermofisher.com account to access theaward-winning, free online Instrument Management Tool*that enables faster responses to requests for service orservice quotes, plus get instant connection to key instrumentand service information.How to reach usGo to thermofisher.com/contactus to find yourlocal support or technical support team. For productFAQs, protocols, training courses, and webinars, go tothermofisher.com/technicalresourcesComprehensive instrument warrantyOur factory-trained and certified field service engineers(FSEs) are focused on delivering the highest-qualityworkmanship. During the warranty period, all repairs,including engineer time and parts, are covered.TaqMan SNP Genotyping AssaysThe TaqMan SNP Genotyping Assays collection includesover 7 million predesigned human and mouse genomewide assays of which 3.5 million are HapMap SNPs,70,000 cSNPs, 160,000 validated assays, and over 2,700drug metabolism genotyping assays. Additionally, ourApplied Biosystems Custom TaqMan SNP GenotypingAssays service lets you create your own custom assays bysubmitting target SNP sequences for any genome.Flexible service plansOur rapid exchange service provides a new or refurbishedinstrument shipped to your lab whenever your instrumentrequires repair. Simply use our prepaid shipping box to returnthe damaged instrument back to us.The AB repair center plan is the cost-effective choice for labsthat send their instrument away for repair—this plan providesan optional loaner instrument so that your lab can maintainproductivity while the instrument is being repaired. Theinstrument is sent back upon repair.**Compliance and validation servicesOur compliance and validation services are designed to helpyou balance business and regulatory requirements. Fromrisk assessment to hardware and software qualification tofull-system validation, we partner with you to help mitigateregulatory risks and get your processes up and running.Training coursesOur application and instrument training programs are ledby scientists who aim to enhance your workday throughexperimental design best practices, workflow training, and* Instrument Management Tool not available in all regions.** Not available in all regions.Reagents and consumablesMeltDoctor HRM Master MixThe Applied Biosystems MeltDoctor HRM Master Mixcontains all components needed for HRM-PCR (excludingtemplate and primers). It is formulated for superior HRMperformance across a wide range of genomic targets. Unlikesome mixes available from other providers, the MeltDoctorHRM Master Mix does not require additional mixing priorto use, and was developed and optimized solely for HRMapplications.RNA isolation kitsA wide range Invitrogen and Applied Biosystems RNAisolation kits are suitable for a variety of sample types,including animal and plant tissue, cultured cells, blood,bacteria, and yeast. Blood, bacteria, and formalin-fixedmaterial require RNA isolation kits designed specifically forthese sample types, whereas most eukaryotic samples canbe processed with excellent results using standard RNAisolation kits.
TaqMan Gene Expression Master MixTailored for quantitative real-time PCR experiments, theApplied Biosystems TaqMan Gene Expression Master Mixdelivers robust performance for both routine and challengingquantitative applications. It enables specific target detectionacross a large dynamic range with high sensitivity down to asingle copy number. Furthermore, it offers duplex capabilityand reproducibility at less than 2-fold discrimination.TaqMan Genotyping Master MixSpecifically formulated for reliable, cost-effective detectionof single nucleotide polymorphism (SNP) detection, theApplied Biosystems TaqMan Genotyping Master Mixprovides accurate and reproducible allelic discrimination withwell-separated clusters for exceptional call rates, premiumperformance in challenging genotyping assays, and excellentbenchtop stability for superior flexibility to meet variousthroughput needs.TaqMan MicroRNA AssaysApplied Biosystems TaqMan MicroRNA Assays canquantitate microRNAs with the sensitivity and specificity ofTaqMan Assay chemistry. Also, the new Applied Biosystems TaqMan Advanced miRNA Assays use ligation-baseduniversal reverse transcription for a streamlined and highlysensitive workflow.Applied Biosystems Primer Express SoftwareApplied Biosystems Primer Express Software v3.0 facilitatesprimer design with TaqMan probes for real-time PCR orendpoint PCR analyses. It is also ideal for primer designusing SYBR Green I dye chemistries. The StepOne andStepOnePlus systems ship with Primer Express Software.For more information about the products, reagents,consumables, assays, and kits listed on this page, go tothermofisher.com/steponeplusTaqMan AssaysWe offer the most comprehensive set of inventoried TaqManGene Expression and SNP Genotyping Assays. Morethan 1.5 million gene expression assays and over 7 millionpredesigned human, and 10,000 predesigned mouseSNP genotyping assays are available at your fingertips.Alternatively, you can submit your target DNA sequencefrom any organism, and we’ll custom-build an assay foryou. TaqMan MicroRNA Assays can quantitate miRNAwith the sensitivity and specificity of Applied Biosystems TaqMan Assay chemistry. For more information on geneexpression assays, go to thermofisher.com/allgenesor for information on SNP Genotyping Assays, go tothermofisher.com/taqmansnpTaqman Gene Expression, SNP Genotyping, and MicroRNA AssaysTaqMan Assays selection guideApplicationGene expression*SNP genotyping†MicroRNA‡TaqMan predesigned Assays(inventoried and made-to-order)YesYesYesCustom TaqMan AssaysYesYesNoSpeciesNumber of inventoried and made-to-order assaysHuman205,0007,000,0005,000Mouse176,000 10,0002,300Rat146,000§1,000Drosophila melanogaster41,000§460Arabidopsis thaliana97,000§340Caenorhabditis elegans92,000§460Canine55,000§N/ARhesus macaque69,000§N/A* Includes miRNA, gene copy number, and mitochondrial assays.† Includes HapMap and drug metabolism genotyping assays.‡ Gene expression only.§ Custom TaqMan Assays are available for any SNP, transcript, and genome.
Reagents and consumablesA complete line of reagents, including TaqMan master mixes,SYBR Green master mixes, and consumables, including96-well plates, is available for use with the StepOne andStepOnePlus Real-Time PCR Systems. These products caneasily be added to a shopping list for future reference or forordering through the materials list link in the experimentaldesign wizard (Figure 11).Figure 11. The reagents ordering page in the experimentaldesign wizard.Select reagents, consumables, and service contract offerings for your StepOne and StepOnePlus Real-TimePCR SystemsCategory/product descriptionQuantityCat. No.MicroAmp 48-Well Optical Adhesive Film25 films4375928MicroAmp 48-Well Optical Adhesive Film100 films4375323MicroAmp 96-Well Optical Adhesive Film25 films4360954MicroAmp 96-Well Optical Adhesive Film100 films4311971Seals and coversReaction platesMicroAmp Fast Optical 48-Well Reaction Plate20 plates4375816MicroAmp Fast Optical 96-Well Reaction Plate with Barcode (0.1 mL)20 plates4346906MicroAmp Fast Optical 96-Well Reaction Plate with Barcode (0.1 mL)200 plates4366932MicroAmp Fast 8-Tube Strip, 0.1 mL125 strips4358293MicroAmp Optical 8-Cap Strip300 strips4323032MicroAmp Fast 48-Well Tray10 trays4375282MicroAmp 96-Well Tray/Retainer Set for VeriFlex Block Systems10 trays43799838-Well stripsAccessoriesMicroAmp 48-Well Base Adaptor5 adaptors4375284Fast SYBR Green Master Mix5 mL4385612Power SYBR Green PCR Master Mix5 mL4367659TaqMan Gene Expression Master Mix5 mL436901610 mL43713552 x 1.25 mL4352042TaqMan RNA-to-CT 1-Step Kit5 mL4392938MeltDoctor HRM Master Mix5 mL4415440ReagentsTaqMan Genotyping Master MixTaqMan Fast Universal PCR Master Mix
Ordering informationDescriptionCat. No.StepOne Real-Time PCR System4376357StepOne Real-Time PCR System with Laptop Computer4376373StepOne Real-Time PCR System with Tower Computer4376374StepOnePlus Real-Time PCR System4376600StepOnePlus Real-Time PCR System with Laptop Computer4376598StepOnePlus Real-Time PCR System with Tower Computer4376599StepOnePlus Real-Time PCR System Upgrade Kit*4379216High Resolution Melt Software v3.0.1 for 1 license4461357High Resolution Melt Software v3.0.1 for 10 licenses4461456StepOne pure dye calibration servicePDC StepOne4460599PDC StepOnePlus4460600Installation qualification and operation qualification (IQ/OQ), operation qualification and instrumentperformance verification (OQ/IPV)StepOne IQ/OQ4413678StepOnePlus IQ/OQ4415138StepOne OQ/IPV4415178StepOnePlus OQ/IPV4415318Service plansStepOne AB Repair Center Support 1 year planZG03SCSTEPONEStepOnePlus AB Repair Center Support 1 year planZG03SCSTEPONEPStepOne Rapid Exchange 1 year planZGEXSCSTEPONEStepOnePlus Rapid Exchange 1 year planZGEXSCSTEPONEPStepOne AB Repair Center Support 3 Year PlanZG03SCSTEPONE3YStepOnePlus AB Repair Center Support 3 Year PlanZG03CSSTEPONEP3YStepOne Rapid Exchange 3 Year PlanZGEXSCSTEPONE3YStepOnePlus Rapid Exchange 3 Year PlanZGEXCSTEPONEP3Y* For users of the StepOne Real-Time PCR System.To learn more about the StepOne and StepOnePlus Real-TimePCR Systems, contact your sales representative, or go tothermofisher.com/steponeplusFor Research Use Only. Not for use in diagnostic procedures. 2016 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiariesunless otherwise specified. TaqMan is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. Excel and PowerPoint are trademarks of Microsoft Corporation.CO020701 0116
PCR functionality and precise temperature control The StepOne system is upgradable to the StepOnePlus system to meet your changing research needs 3 StepOnePlus Real-Time PCR System Upgrade The Applied Biosystems StepOnePlus Real-Time PCR System Upgrade is designed to accommodate your changing real-time PCR needs. The upgrade converts a
