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SPECIFICATION SHEETStepOne and StepOnePlus Real-Time PCR SystemsApplied Biosystems StepOne andStepOnePlus Real-Time PCR SystemsRemarkably Simple Systems. Simply Remarkable Results.Total ConfidenceStepOne and StepOnePlus Systems Data stored both on the instrument andcomputer for extra safetyStepOne System Fast system with plug and play assays andfast reagents for maximum time savings Ultra-compact footprint fits mostlaboratory settings48 wells for lower throughput3 colors for basic applications researchEasily upgradeable to the StepOnePlus SystemStepOnePlus System96 wells for higher throughput4 colors for more flexibilityVeriFlex Blocks technology forthermal cycling flexibility LCD touchscreen and USB drive with theoption of PC‑free instrument operation Remote monitoring and email notificationwith run file attached for immediate accessto results–even remotelyThe Applied Biosystems StepOne and StepOnePlus Real-Time PCRSoftware Highlightsa user-friendly interface. Flexible and easy to use, these systems can be set Intuitive, flexible software with plate set upwizard that guides new users through theirreal‑time PCR experiments Powerful Gene Expression Study Packagewith advanced features for complex studiesSystems are remarkably powerful real-time PCR instruments designed withup and operated with total confidence, even by researchers who have little orno previous real-time PCR experience. Highly affordable, these new low- tomedium-throughput systems are suitable for every level of experience. Theeasy to use software with its optional wizards guides researchers through allaspects of sample processing, including sample and reaction setup, thermal Customizable graphs and plots Export to JPEG, Excel and PowerPointcycling, and fluorescent detection. The StepOne and StepOnePlus Systems Optional software package offers theability to conduct high resolution melt(HRM) experimentseven assist users in selecting and ordering real-time PCR reagents online.

Table 1. Features at a Glance.StepOne SystemStepOnePlus SystemThroughput/Wells4896FAM /SYBR Green dyes VIC /JOE dyes ROX dyes NED/TAMRA dyes Veriflex Blocks Plug and play applicationsStepOne Software Designed for New andAdvanced UsersInstrument software for the AppliedBiosystems StepOne and StepOnePlus Real-Time PCR Systems provides instrumentcontrol, data collection, and data analysis.This user-friendly and intuitive softwarepackage includes the following features: Experimental Design Wizards to help youdesign and set up experiments Advanced setup for expert users whorequire more flexibility for more complexapplications, such as multiplexing QuickStart setup so runs can beginimmediately and plate information enteredlater Remote real-time monitoring for observingthe run from a remote PC Email notifications with data file attached,providing immediate access to results Exports data to PowerPoint , Excel , or.jpeg files Troubleshooting flags to help diagnose andsolve problematic experiments Multiple Plots view for simultaneous dataassessment from four perspectivesPrimer Express Software v3.0 is alsoincluded with the systems. This softwareoffers simple primer and probe design forreal-time PCR applications. Powerful Gene Expression Study PackageImport an unlimited number of ComparativeCt (Relative Quantitation) plates to one study: View data by biological replicate group ortechnical replicate group Normalize to multiple endogenouscontrols Enter known efficiencies to adjust RQvalues for each target View amplification plot, multi-componentplot and QC summary within the study toeasily identify and eliminate outliers Preview the effect of modified analysissettings before permanently applying themto resultsHRM Just Got A Lot EasierOptional High Resolution Melt Software v3.0offers the ability to conduct high resolutionmelt experiments on the StepOne andStepOnePlus Systems: Accurately distinguish control and variantgenotypes with a new clustering algorithm Input expected number of clusters fordifficult-to-detect SNPs Run separate analysis of multiple assaysrun on a single plate Copy and paste directly from Excel software Easily view well information and platelayout in the Plate View Saves analysis settings for futureexperiments—software automaticallyapplies default/saved settings uponopening an experiment file As Flexible as Possible with PC-Free andNetworkable OperationThe StepOne and StepOnePlus Systemscan be installed in five distinct configations:1. PC controlled2. PC free3. Networked4. PC controlled connected to LAN5. PC controlled with network instrumentConnection to an ethernet network enablesremote monitoring, downloading and uploading of experimental files and data.StepOne and StepOnePlus Systems areavailable for purchase with either a laptop ortower computer. Systems without computersare also available. The instruments can beoperated even without a PC—a QuickStartfeature initiates experimental runs from theinstruments touchscreen.Convenient Online OrderingFor convenient ordering of real-time PCRmaster mixes and consumable plasticware,an experimental material list is accessiblethrough the Design Wizard in the systemsoftware. It links directly to the AppliedBiosystems Store (optional).

Table 2. Demonstrated Performance Specifications.Dynamic Range9 logs of linear dynamic rangeSensitivityA single-reporter TaqMan Assay can detect 10 copies of RNaseP in a 30 μL reaction volumePrecisionUsing the TaqMan RNaseP Instrument Verification Plate, the system can distinguish between 5,000 and 10,000template copies of RNaseP with up to 99.7% confidenceRun TimeFast: 40 min/40-cycle PCR reactions, using the TaqMan RNaseP Verification Test Plate Standard: 2 hours/40cycle PCR reactionsFluorescence DetectionFluorophore detection chemistries includeFAM , VIC /JOE and ROX dye-labeledTaqMan MGB probe-based assays. Withthe StepOnePlus System, TAMRA dye canalso be utilized. The fluorophore detectionchemistries offer outstanding specificityand sensitivity for real-time quantitation,gene expression assays, genotyping assays,and multiplex reactions. The systems alsoaccommodate SYBR Green I dye assaychemistry, an economical alternativefor target identification, initial screeningassays, or for studies that require only a fewreactions.All sample wells in these systems areilluminated with a high powered blue LEDexcitation light that is designed to last forup to ten years of continuous use, virtuallyeliminating the need for replacement.Fluorescence emission is detected throughfilters on a photodiode. The emission filtersare optimized for use with FAM /SYBR Green I, VIC /JOE , and ROX fluorescentdyes.Experiment Ready on Day OnePrior to shipping, each system is calibratedfor optical and thermal accuracy. During theinstallation setup, which requires 4 hours,the RNaseP Instrument Verification TestPlate confirms performance specifications tohelp ensure data fidelity.Innovative VeriFlex BlocksThe 96-well StepOnePlus System featuresVeriFlex technology, which brings sixindependently controllable peltier blockstogether for precise temperature control andenhanced PCR functionality. VeriFlex blocksdeliver flexibility for those who have the needto optimize their assays or who have probesand primers that are optimized at differentannealing temperatures. The VeriFlex blocks can run with six different annealingtemperatures in the same run.StepOnePlus Real-Time PCR SystemUpgrade KitTo accommodate your changing real-timePCR needs, Applied Biosystems offers forsale the StepOnePlus Real-Time PCRSystem Upgrade Kit. The upgrade convertsa StepOne System into a StepOnePlus System by upgrading the 48-well block toa 96-well VeriFlex Block and expandingthe dye flexibility to four colors. The colorof the bar above the block is also updatedto indicate that the instrument is now aStepOnePlus System. Just send in yourexisting StepOne System and you’ll receivea StepOnePlus loaner instrument until yourupgraded system arrives in your lab.Service and WarrantyPurchase of the StepOne or StepOnePlus Real-Time PCR System includes a limitedwarranty* on parts and labor. AppliedBiosystems provides worldwide technicalsupport and service. Depot repair servicesare available for both systems throughApplied Biosystems.Validated SolutionIQ/OQ - A certified Applied BiosystemsService Engineer will assist with theInstallation Qualification (IQ/OQ) processas part of the overall system validation.According to your change control and maintenance requirements, they can re-Qualify(OQ/IPV) to the same specifications; verifyingconsistent performance over time. Theseservices are available for a separate fee.*Warranty length varies by region.

Table 3. Instrument Specifications.StepOne SystemStepOnePlus SystemThermal cycling systemPeltier-based systemPeltier-based systemBlock format48-well block96-well blockSupported volumes10–30 μL10–30 μLSupported consumables 48-well (0.1 mL) plates with optical adhesive covers 96-well (0.1 mL) plates with optical adhesive covers 48-well (0.1 mL) plates with optical flat caps 96-well (0.1 mL) plates with optical flat caps 8-tube (0.1 mL) strips with optical flat caps 8-tube (0.1 mL) strips with optical flat caps Individual (0.1 mL) tubes with optical flat caps Individual (0.1 mL) tubes with optical flat capsSample ramp rateFast mode: /- 2.2 C/secStandard mode: /- 1.6 C/secFast mode: /- 2.2 C/secStandard mode: /- 1.6 C/secPeak block ramp rate4.6 C/sec4.6 C/secVeriFlex BlocksN/A25 C (5 C zone-to-zone)Temperature range4 C–100 C4 C–100 CTemperature accuracy 0.25 C (35 C to 95 C) of setpoint/display temperature,measured 3 minutes after clock start 0.25 C (35 C to 95 C) of setpoint/display temperature,measured 3 minutes after clock startTemperature uniformity 0.50 C, measured 30 seconds after clock start overthe temperature range of 35 C to 95 C 0.50 C, measured 30 seconds after clock start overthe temperature range of 35 C to 95 CMelt curve resolutionAs small as 0.1 CAs small as 0.1 COptical systemLED, emission filters, photodiodeLED, emission filters, photodiodeCalibrated dyes at installationFAM , SYBR Green I, VIC , JOE , ROX dyesFAM , SYBR Green I, VIC , JOE , NED, TAMRA, ROX dyesPassive reference dyesROX dyeROX dyeData collectionData collected in all filters for all wells regardless ofplate setup. Plate setup may be modified after runcompletesData collected in all filters for all wells regardless ofplate setup. Plate setup may be modified after runcompletesQuantitative PCR Run TimeFast: 40 minutesStandard: 2 hoursFast: 40 minutesStandard: 2 hoursTouchscreenLCD/6.5 in. VGA (640 x 480)/32K colorsLCD/6.5 in. VGA (640 x 480)/32K colorsInstrument dimensionsWidth: 24.6 cm (9.7 in)Depth: 48.5 cm (19.1 in)Height: 51.2 cm (20.4 in)Weight: 23.6 kg (52 lbs)Width: 24.6 cm (9.7 in)Depth: 48.5 cm (19.1 in)Height: 51.2 cm (20.4 in)Weight: 24 kg (53 lbs)Software SpecificationsApplications—Comparative Ct, Standard Curve, Relative Standard Curve, Genotyping Presence/Absence, MeltCurve Analysis, high-resolution melt (HRM) analysis (additional software package required)Dye Discrimination—Multicomponenting algorithmMultiplate Data Comparison—Compare an unlimited number of plates of gene expression assaysMultiplex Capability—Multiplex up to five targets per wellPC Compatible