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AriaMxReal-Time PCR System
TOTAL CONFIDENCE qPCRBuilt on Stratagene’s legacy of enzyme engineering expertise and precision instruments, AgilentTechnologies provides a comprehensive approach to real-time quantitative PCR (qPCR)—fromsample preparation to data analysis. Every Agilent qPCR instrument, including the AriaMx Real-TimePCR System, comes with the same quality, intuitive software, and technical support that you havecome to expect from the successful Mx3000P and Mx3005P qPCR instruments on the markettoday. We are proud to carry forward the people and product excellence from the Stratagene familyto the Agilent family—ready to support you at every step.Whether you are new or experienced in qPCR, Agilent’s full range of products and industry-leadingsupport is sure to keep you up and running with complete confidence and superior results.SPEED XPERTISE2
SPEEDConfidence in Success with BrilliantUltra-Fast Master MixesOur unique Brilliant reagents are equally robust and reproducible across avariety of assays—even on fast cycling platforms. Superior specificity isdelivered utilizing a novel, faster-activating hot start method to minimize theformation of primer-dimers and other off-target reactions. Enhanced rapid hot start capability Reliable and reproducible dataAGILITYConfidence in a System that MeetsYour Needs—Today and TomorrowThe agile design of the AriaMx Real-Time PCR System offers the industry’sfirst configurable and customer-changeable optics. Now you can mix-andmatch optics to suit your needs today, and easily change them to suit yourneeds for tomorrow. Modular design Ready-to-use Future-proofPRECISIONConfidence in Easy-to-Use,Precise SoftwareThe AriaMx Software combines robust data analysis algorithms andintuitive organization for precision and ultimate ease-of-use. Control experimental bias due to differences inamplification efficiencies through a proprietary algorithm Multiple, customizable data analysis algorithms Thermocycling control and precision you expect froma more expensive instrument 21 CFR part 11 compliant software3
AriaMxBETTER SYSTEMS. BRILLIANT RESULTS.Confidence in a system that meets your needs today and tomorrow.The AriaMx Real-Time PCR System is a fully integrated qPCR amplification,detection, and data analysis system. The system’s modular design combinesa state-of-the-art thermal cycler, an advanced optical system with spectraoptimized LED cartridges, and data analysis software.The instrument leverages a comprehensive software suite of on-board instrumentdiagnostics, giving you confidence that instrument failpoints are identified prior torunning your assay. Experience total confidence with AriaMx’s blend of speed, agility,and precision.PRE-RUN TEST: 60 ATTRIBUTESIN 1MIN4PRECISETHERMALUNIFORMITY
SPEEDAGILITYFactory CalibratedAriaMx is ready to go—no need to setquantification curves or calibratethe instrument.Modular DesignModular cartridges make in-lab upgradespossible with the click of a button.Ready, Go!Today and TomorrowUltra-Fast ChemistryAriaMx is optimized for Agilent’sBrilliant III reagents, but also runs all fastchemistries.Blazing Scan TimesScan all channels in less than threeseconds—the fastest in the industry.Intuitive InterfaceAriaMx’s touchscreen makes systemintegration easy for all users.Broad ApplicationsAriaMx supports more than one assay andchemistry. Applications include: GeneExpression, Comparative Quantification(Single-Plex and Multiplex), QuantitativePCR (Single-Plex and Multiplex),Genotyping, Allele Discrimination, NGSLibrary Quantification, and more.PRECISIONSuperior DataThermal ControlEasily maintains within 0.2 C orless of target temperature.Impressive SensitivityTwo-fold discrimination in a singlecycle with 95% confidence over awide range of copies.Robust ValidationTested on 100,000 samples.MODULAROPTICSEASY-TOLEARN5
SPEEDBRILLIANT REAGENTS Built upon Stratagene’s legacy,Agilent Technologies provides a total solution approachto real-time quantitative PCR.Ultra-Fast SYBR Master Mixes Robust validationand tightmanufacturingquality controlsA proprietary, quick acting hot start Taq mutant enables ultra-fastreactions which maintain amplification efficiency, R2, dynamic rangeand detection sensitivity. SYBR assay resultsin 42 minutes usingBrilliant III reagents Open reagentplatform: use yourassays and reagents AffinityScript for thetoughest samplesand alleles6SPEED. AGILITY. PRECISION.
AriaMx instrumentation makes routineand complicated applications faster andeasier without sacrificing data integrity.Applications where AriaMx excels: m iRNA QuantificationMaster mixes which enable yourassays to discriminate miRNAdifferences—even by one SNP HRMConfidently genotype the toughestalleles in less time Open PlatformInstrumentation validated for allmajor manufacturer kits as wellas Agilent reagents MultiplexMore agility and speed in yourassays with reagents optimized foryour reactions N GS Library QuantificationOptimal cluster density improvingefficiency and data qualityFIG. 1 Six Color Standard Curve Data by FluorGREATERRESISTANCE TOINHIBITORSFIG. 2 SYBR Green Assay—Amplification and Standard Curve Plots Showing Sensitivity Measuring a Two-Fold ChangeThe standard curve consisted of nine, ten-fold dilutions starting with 1x109 copies and ending at two copies. Amplification plots are below.7
SPEEDBRILLIANT HRM ULTRA-FAST LOCI MASTER MIXFast chemistry to confidently identifyhard-to-detect genotypes.High ResolutionMelt ApplicationHigh resolution melt analysis (HRM)is a quick method to monitor andrecord the melt profile of ampliconsin a sample, post-PCR. HRM’s highsensitivity can detect even thesmallest of melting temperaturechanges like those caused by singlebase changes, nucleotide repeatsand small deletions in DNA.Some of the more commonapplications of HRM include:genotype confirmation, mutationidentification and screening, cloneconfirmation and methylationanalysis.HRM is often used at hypervariableloci to find hard-to-detect basechanges. In contrast to TaqManassays, which can easily miss newor unknown mutations, HRM candetect any mutation between twoprimers in the assay. HRM alsocost-effectively resolves smallchanges while maintaining meltingtemperature control.8Brilliant HRM UltraFast Loci Master MixAdvantagesHigh-Performance Agilent HRM Options: M ix and GoMgCl2 and dNTPs are included. B etter stabilityStable after multiple freeze thaws,reducing wastage and increasingbatch-to-batch reliability. Fast-Start TaqThree-minute activation time forthe proprietary mutant Fast-StartTaq polymerase.Brilliant HRM Ultra-Fast Loci Master MixFaster HRM on any platform with total confidence EvaGreen Release-on-Demandbased dyeNon-toxic and can be addedat saturating concentrations—insuring minimal inhibition whilepreserving high sensitivity. P latform independentUse with any HRM-capablethermocycler.Complete AriaMx HRM SolutionThe fastest way to confidently identify hard-to-detectgenotypesSPEED. AGILITY. PRECISION.
Agilent HRM Ultra-Fast Loci Master MixFor the scientist looking to quickly “mix-and-go” with greater confidence, Agilent now offers the Brilliant Ultra-Fast Loci Master Mix asa stand alone reagent—validated to work on your existing instrumentation. Although optimized for Agilent’s AriaMx, the master mixalso works on most major manufacturer instrumentation.The HRM Ultra-Fast Loci Master Mix combines a fast-start mutant Taq polymerase, MgCl2 buffer optimized for most HRM reactions,dNTPs and an EvaGreen low toxicity dye, considered the most reliable for HRM.FIG. 3 HRM of SNP Rs9939609 (FTO) Using Brilliant HRM Master Mix on Three Different InstrumentsPanel A (Agilent AriaMx qPCR System)SNP identified using Brilliant HRM Master Mix onAgilent’s AriaMx qPCR Instrument.Panel B (Competitor B)The same SNP identified using Brilliant HRM MasterMix on Competitor B qPCR Instrument.Panel AB (Competitor AB)The same SNP identified using Brilliant HRM MasterMix on the Competitor AB qPCR Instrument.9
SPEEDBRILLIANT HRM ULTRA-FAST LOCI MASTER MIXComplete HRM Solution for total confidencein your data.Intuitive HRM SoftwareAgilent includes fully featuredsoftware for HRM analysis whichcontains several benefits over othersoftware packages with respect toplate set up, analysis and reporting(Table 1). Unlike competitive HRMsoftware, Agilent HRM softwarecomes standard with the AriaMxReal-Time PCR System—includedwith the instrument at no charge.The Agilent platform’s speedadvantage over the competitionis due to fast scan times and analgorithm used to interpolate themaximum Tm peak value. Otherplatforms report the highest meltpeak value observed during the run,requiring additional scans to obtainan equivalent answer.10Table 1. Comparison of Instrument Software for HRM AnalysisFeatureInstrumentA(AriaMx)BABSet up replicates quickly using “smart-rulesbased” softwareUse files from a previously-run experimentas a templateUse a highlighting feature to link plots tothe results table for easier analysisHover over each plot to reveal key data(e.g. X, Y coordinates, replicate number,dye channel)YesNoPartialPart NumberDescription5190-7827Brilliant III HRM Ultra-Fast Loci Master Mix5190-7702AriaMx HRM Calibration KitSPEED. AGILITY. PRECISION.
Agilent Complete HRM SolutionFor researchers ready for a change in genotyping speed and confidence, Agilent offers a complete solution: HRM Master Mix, aninstrument and analysis software.FIG. 4 Agilent AriaMx qPCR Instrument is 20% Faster than Competitor InstrumentsHot Start69A: UFLThis graph shows the time required to identify the Class IV SNPRs9939609 FTO with the Brilliant HRM Ultra-Fast Loci Master Mix(UFL) on three different qPCR instruments. The assay consisted of ahot start, 40 cycles of qPCR amplification and a high resolution meltsegment. The time differences observed are due to: 1) differences ininstrument scanning speeds, 2) the number of measurements overthe temperature profile, and 3) Taq polymerase’s activation time.AmplificationHRM Segment88B: UFL108AB: UFL050Time (minutes)100150A: AriaMx instrument with Brilliant HRM Master MixB: Competitor B qPCR instrument with Brilliant HRM Master MixAB: Competitor AB qPCR instrument with Brilliant HRM Master MixFIG. 5 Agilent Complete HRM Solution is 30% Faster than Competing HRM SolutionsHot StartA69This graph shows the time to identify the Class IV SNP Rs9939609FTO on three competing qPCR instruments with manufacturer’srespective HRM chemistries. Manufacturer’s recommended HRMprotocols were used.AmplificationHRM Segment101B185AB050Time (minutes)100150A: Agilent AriaMx qPCR Instrument with Agilent Brilliant HRMUltra-Fast Loci Master MixB: Competitor B qPCR instrument and competitor B’s HRMMaster MixAB: Competitor AB qPCR instrument with competitor AB’s HRMMaster MixUsing Agilent’s complete HRM solution, the time to correctly identify the most difficult-to-detect genotypes can be cut in half(Figure 5). The longest run time (185 minutes) was on the AB instrument with the AB reagents. In contrast, on the Agilent AriaMxinstrument with the Brilliant HRM Ultra-Fast Loci Master Mix, the same result was obtained in only 69 minutes. Now researcherscan identify difficult-to-detect genotypes much faster than any other 96-well plate based solution—with ease and confidence.11
AGILITYMODULAR OPTICAL CARTRIDGES The first real-timePCR system with optics contained in modular cartridges,the AriaMx allows for less cross talk between channels—resulting in improved data resolution.MULTIPLEXFROM 2 TO 6CHANNELSReady, Go!Key FeaturesAriaMx comes calibrated straightfrom the factory. Calibration is onlynecessary if future modules areadded. Now you can be confident youare measuring biological variationover the instrument’s life. Expandable and upgradeable in the lab No reference channel needed “Set it and forget it” calibration Future-proof A ccommodate as many or as few opticchannels as you requireWith optical cartridges that have noneed for a reference channel, thetight calibration of the cartridges andsteady emission wavelengths from theinstrumentation provide consistentdata with every run.12SPEED. AGILITY. PRECISION.
Fig. 6 Validation Run With No ROX Reference(Above) and With ROX Reference (Below)*AGILITYUPGRADE YOURTIMETABLEWhen detecting biologically relevant changes,which occur at low thresholds, instrumentsensitivity is critical. The SYBR uniformity curvesat left were generated as part of the AriaMxvalidation on the same day. Both begin at cycleseven and end at cycle 17.* Sigma multiplier was set to 19 for both runs.Adaptive smoothing was also on for the baselinegraph. dR, SD 0.13 Cq for both runs.Better multiplexing: Don’t sacrifice time, samples or reagentsWe know you’re always looking tomaximize information obtained from raresamples. Multiplexing is one solution togetting answers in the least amount oftime. However, multiplexing often requiresextensive optimization of reagents, whichcan be expensive in both time and reagentcosts and is quickly becoming animpractical method. Agilent’s BrilliantMaster Mixes allow you to normalize yourresults with internal controls, providingthe most accurate quantification possible.Agilent Brilliant Multiplex qPCRMaster MixesProviding sensitive, real-timeamplification, Agilent Brilliant MultiplexqPCR Master Mix allows the use ofinternal controls to provide normalizationwithin each reaction while reducingtime and reagent costs. The BrilliantMultiplex qPCR Master Mix providesamplification of up to four targets perreaction, and each requires far lesstemplate than if performed in fourseparate reactions. Moreover, thesensitivity remains equivalent to thatseen in single-plex.Agilent Complete Multiplex SolutionAriaMx delivers answers frommultiplexing experiments faster than mostthermocycler-based platforms. Get readyfor an improvement in time-to-results andaccuracy.Part NumberDescription600880Brilliant III Ultra-Fast QPCR Master Mix for Multiplex Reactions600884Brilliant III Ultra-Fast QPCR Master Mix for Multiplex Reactions13
PRECISIONAdvanced, yet easy-to-use SOFTWARE with proprietaryalgorithms accompany AriaMx’s precision chemistry andthermocycling capability.Fig. 7 Easy-to-access Plate and Well MapsEasy Touchscreen Set-UpPlate Maps at Your FingertipsSimple-to-use, feature-rich touchscreenallows you to set up runs with the touchof a finger.Program at the plate and well level. Simplyhover over a well, and its details will openin a new window.21 CFR Part 11 compatible featuresare available by licensing ET software.New Software Features Touchscreen Set-Up Onboard Diagnostics Remote Monitoring Base software included withinstrument purchase14SPEED. AGILITY. PRECISION.
Fig. 8 Set-up Screen for Comparative QuantitationEasy ReportingEasily export raw data in multipleformats with the touch of a finger.Select only those datasets you wantdisplayed using custom data reports: Plate Set-Up Experimental Thermal Profile Raw Data Plots Analysis Criteria Amplification Plots Allele Determination Graphical DisplaysAriaMx Software helps you control experimental bias due to differences in amplificationefficiencies through a proprietary algorithm—resulting in higher accuracy.Pre-programmed AssaysMultiple, CustomizableData Analysis Algorithms Tabular Results Experimental NotesFig. 10 Report ConfigurationPre-programmed assays allow foreasy selection of calibrators,Identify NGS Bias and Fragment Dropnormalizers and sample associations.Accurate NGS library quantificationis crucial for determining sequencing Quantitative PCRefficiency and data quality. Allele DiscriminationCombine the AriaMx System with the Comparative QuantificationAgilent Bioanalyzer for a powerful gene High-Resolution Meltexpression and next-gen sequencingIn high throughput gene expression solution. See www.agilent.com/studies, standard curves are notgenomics/bioanalyzer for more details.used. Relative quantity calculation isautomated using the ComparativeQuantitation module in the AriaMxSoftware. See Figure 8 for details.Fig. 9 Relative Gene Expression15
SERVICESGive your research the attention it deserves. Regain timelost on peripheral tasks and focus on the critical processesthat keep your lab competitive and successful. Speedy delivery forcritical parts Dedicated instrumentsupport for fast issueresolutionMore Reliable Resultsand AchievementsAgilent service agreements, preventativemaintenance, and compliance services aresold to you at the system level, assuringall modules of your system are covered.Confidence, Not CompromiseAgilent’s service group stands behindevery instrument, giving you confidence inequipment functionality and data. Agilenthas one global focal point: providing ourcustomers with the best service availableworldwide. It’s no wonder that over70% of our service employees stay withAgilent for over five years. Equipped withcalibrated and traceable tools, our servicetechnicians verify your equipment isrunning at maximum performance. Agilenthardware and software qualificationprotocols ensure rapid conflict resolution.Specialized remote monitoring (free toactivate as needed) allows Agilent’s servicegroup to troubleshoot your instrumentdirectly from the lab, providing confidenceat every turn.Breadth of ServicesAgilent’s professional services teamhas been called upon for our supportwith customized training, systems andenterprise integration, workflow design,special engineering projects, protocoldevelopment, and project management.We have conducted over 100,000successful system qualificationsworldwide, and our genomicsprofessionals operate in 65 countries.Rapid AssistanceWhatever the challenge, Agilent’s broadresources address your needs. Expectreduced repair time with: Call Center: Rapid assistance withinstrumentation, reagents, applications,and protocols requires a vastknowledge of a variety of chemistries.That’s why our technical supportprofessionals are PhD scientists withgenomics experience—just like you. Detailed Asset Information: Agilent’stracking system provides criticalinformation on instrument usage,resulting in quicker solutions. Wireless Technology: Enables ourservice team to provide instant accessto a database of information andresources. Remote Monitoring: Instrumenthealth monitored remotely for greatertroubleshooting flexibility.16
Warranty and Multi-YearUpfront CoverageEach AriaMx has a global warranty,including the standard warranty forthe country of purchase. If moved toanother country, the destinationcountry’s standard warranty willapply to the instrument.*Services available for purchase withAriaMx include: Return to Agilent (RTA): This may beaccomplished with a loaner instrument,exchanging out your instrument, or directreturn of your instrument to Agilent. Installation and Familiarization (I&F):Have an Agilent Field Service engineeror member of our team install andfamiliarize you with the AriaMx. Standard Preventative Maintenance(PM): Agilent is available to run routinemaintenance check-ups on yourinstrument annually to ensure peakperformance for the upcoming year.**COSTEFFECTIVEON-TIMEDELIVERY Extended Preventative Maintenance:Conducted annually on your instrumentand includes thermal block tests toevaluate uniformityand precision.** Silver Service Package: Bundledservice including Standard PreventativeMaintenance check-ups and the Returnto Agilent (RTA) program, if necessary.1-year base warranty sold with the instrument. Additional coverage can be purchased at a minimal cost.1-YEARInitial WarrantySold with instrument(covers 1 year) 1 YEARExtended Warranty 2Adds additional year(covers 2 years) 2 YEARSExtended Warranty 3Adds additional year(covers 3 years) 3 YEARSExtended Warranty 4Adds additional year(covers up to 4 years) 4 YEARSExtended Warranty 5Adds additional year(covers up to 5 years)* Except for on-site warranty where Agilent does not have an applicable product-specific supportpresence or authorized representative in that country.** Unless a service contract is in place, parts replaced outside the scope of preventative maintenancewill be charged as a repair event (this includes the engineer’s time and materials).17
Excitation Source8 dye specific LEDs per optical moduleSample Containers96-well plates, strip tubes; 0.2 mL tubesDetection Sources8 photodiodesWarrantyOptical CartridgesSYBR/FAMHEXROXCY3CY5ATTO4256 slots, swappable optical modules 1-year warranty is standard with the instrument 5-year warranty and service packages availableOnboard Analytics Thermal, physical, interactive (sensors) tests Extended: 125 performance points tested in30 minutes Start-up: 59 performance points tested in 1 minute Optional bypass of both featuresServices (upon request) Installation and familiarization Standard and Enhanced Preventative Maintenance Additional year warranty ( 1 increments, up to 5years coverage) Return-to-Agilent Instrument Exchange Program Thermal block verificationOperating System Windows 7 and 10MS Office Compatibility Microsoft 2010 and 2013 compatibleRun Modes Stand alone PC connected L AN connected to PC (more than 20 instrumentscan be connected and monitored remotely) USB connected, external devicesSoftwareFree software including LIMS connectivityOptical ModuleCalibration andCleaning All channels can be tested and calibrated A ll attributes of optical channels are calibratedat the factory – LED light output, light path, mirror,and photodiode Optical modules can be cleaned in lab withoutAgilent technician or sending back to factorySelected Applications Quantitative and qualitative gene expressionanalysis miRNA analysis Genetic mapping Genetic fingerprinting NGS library quantification 2-6 channel multiplex ability HRM analysis (including genotyping, mutationalanalysis, and class IV SNP detection) Pathogen quantificationDye SelectionExcitation and EmissionReaction Volume10 μL to 30 μLChemistries SupportedSYBR, Probe, HRMThermal SystemSix Peltiers made from two ceramic plates withsemi-conductor elements, 96-wellThermal SystemTemperature Range25.0 – 99.9 CHeating: 6.0 C/secCooling: 3.0 C/sec (Median), 2.5 C/sec (Average)Accuracy: 0.2 C or better at typical annealing,amplification, and denaturation temperaturesDynamic Range9Experiment TypesQuantitative PCR with dye, Quantitative PCRwith probe, Allele Discrimination with HRM,Allele Discrimination with probe, ComparativeQuantitation, User DefinedUniformity 0.4 CData Acquisition Time 3 seconds for allCq UniformityCq St Dev 0.20 at fast cycling (5s 95 C/10s 60 C)Electrical Power (input)100 – 240VAC, 50/60Hz, 1100VAOperating Environment20 – 30 C, 20 – 80% non-condensing humidity,7500 feet, max altitudeWeight50 lbs. (23 kg)Dimensions19.7” W x 18.1“ D x 16.5” H(50cm x 46cm x 42cm)18
SPEEDAGILITYPRECISIONQuick ReferenceUseful ToolsqPCR Decision Tree: Determine which qPCR reagents or enzymes best serve your needs.Software Updates: Ensure your software is the most current version.qPCR Methods and Applications Guide: Useful for both new and expert users.Field Notes: Updates and quick tips on AriaMx, from new applications to protocols you can rely on.19
Ordering InformationCategoryPart NumberDescriptionQuantityBase InstrumentationG8830AAriaMx Real-Time PCR System1Optical Cartridges(*Select 1-6 filters to add to baseinstrument. Additional filters canbe added to base instrument atany time)G8830-67001*SYBR/FAM Optical Cartridge1/packG8830-67002*ROX Optical Cartridge1/packG8830-67003*HEX Optical Cartridge1/packG8830-67004*CY3 Optical Cartridge1/packG8830-67005*CY5 Optical Cartridge1/packG8830-67006*ATTO425 Optical Cartridge1/packG5380AAElectronic Tracking of HRM software1G5381AAHRM analysis software upgrade1G4983AAAriaMx Software on HP Laptop1401490AriaMx 96-well plates, skirted and low profile1 x 25/pack401491AriaMx 96-well plates, skirted and rigid1 x 25/pack401494AriaMx 96-well plates, non skirted low profile1 x 25/pack401492AriaMx adhesive plate seals1 x 50 plates401493AriaMx low profile strip tubes for PCR and qPCR applications,without caps8/strip x 120/box401425AriaMx Strip caps for PCR and qPCR applications8/strip x 120/box600880Brilliant III Ultra-Fast QPCR Master Mix400 rxns600881Brilliant III Ultra-Fast QPCR Master Mix, 10 pack10 x 400 rxns600882Brilliant III Ultra-Fast SYBR Green QPCR Master Mix400 rxns600883Brilliant III Ultra-Fast SYBR Green QPCR Master Mix, 10 pack10 x 400 rxns600884Brilliant III Ultra-Fast QRT-PCR Master Mix400 rxns600885Brilliant III Ultra-Fast QRT-PCR Master Mix, 10 pack10 x 400 rxns600886Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix400 rxns600887Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix, 10 pack10 x 400 rxns302106MycoSensor QPCR Assay Kit100 rxns302107MycoSensor QPCR Assay Kit50 rxns5190-7708AriaMx SYBR Qualification Plate1 plate/pack600553Brilliant Multiplex qPCR Master Mix200 rxns5190-7827Brilliant HRM Ultra-Fast Loci Master Mix200 rxns5190-7702AriaMx HRM Calibration Kit1 x 96-well plateStarter Packs600906*AriaMx SYBR Green Starter Pack1(*Includes cDNA synthesis kitsand recommended plastics)600907*AriaMx qRT-PCR Starter Pack15190-9370Brilliant HRM Ultra fast Starter Pack1Software Upgrade and PCPlasticsReagentsPROBESYBRMULTIPLEXHRMFor Research Use Only. Not for use in diagnostic procedures.PR7000-0129 Agilent Technologies, Inc. 2020Published in the USA, October 20, 20205991-9159EN
Ultr a-Fast Master Mixes Our unique Brilliant reagents are equally robust and reproducible across a variety of assays—even on fast cycling platforms. Superior specificity is delivered utilizing a novel, faster-activating hot start method to minimize the formation of primer-dimers and other off-target reactions.
