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molecular biologyessentialsTaqMan is a registered trademark of Roche Molecular Systems, Inc. Used under permission and license.

you may know usbetter than you thinkLook closer—your favorite life sciences products are now by Life TechnologiesFor years, you’ve trusted our products and services for your everyday life sciences needs.As Life Technologies, we’re delivering that same quality in a way that’s simple. Today, onepartner for scientific innovation. Soon, one source for ordering, service, and support. We’reone company driven to help you advance discovery.www.lifetechnologies.com/one 2011 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners, unless otherwise noted.TaqMan is a registered trademark of Roche Molecular Systems, Inc. Used under permission and license.

real-time PCRnucleic acid purification and analysisend-point PCRcloningRNAi, epigenetics, and non-coding RNA researchprotein expression, isolation, and analysisArcturusXT laser capture microdissection system1234567I

introduction2011/12 Life TechnologiesMolecular Biology Essentials CatalogueintroductionComprising trusted and familiar Invitrogen and Applied Biosystems products, the Life Technologies molecular biologyportfolio offers life sciences researchers the most-published, high-performance, scalable solutions across the breadth ofmolecular biology applications.The chart below shows an organized snapshot of these solutions, which have been cited in more than 200,000 publicationswithin the past 10 years. To help you quickly find products that meet your needs, we’ve included icons to denote benefits liketime-savings and high-throughput. Refer to the chapter noted for complete product information.Gene analysis and cloningSamplepreparation Keytechnologies Detectioninstruments Essentials Time saverIIPureLink Plasmid Prep (Ch. 2)PureLink RNA Mini Kit (Ch. 2)BenchPro 2100 Plasmid PurificationSystem (Ch. 2)MagMax sample preparation system (Ch. 2)iPrep Purification Instrument (Ch. 2) GeneArt Gene Synthesis and Gene Assembly(Ch. 4) Invitrogen cloning Protein expressionSuperScript Reverse Transcriptase, TOPO cloning, Gateway, TA Cloning (Ch. 4)Platinum Taq, AmpliTaq Gold 360 MasterMix (Ch. 3)E-Gel DNA & RNA separation gels (Ch. 2) Invitrogen Qubit DNA & RNA QuantitationSystem (Ch. 2)Applied Biosystems Veriti , 9700,2720 thermal cyclers (Ch. 3) TrackIt DNA Ladders (Ch. 2)Oligos/primers (Ch. 3), dNTPs (Ch. 1),Invitrogen One Shot competent cells (Ch. 4),UltraPure agarose & reagents (Ch. 2)Ambion RNA Essentials (Ch. 2), ligases& restriction enzymes (Ch. 4) High throughputFor Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. 2011 Life Technologies Corporation. All rights reserved.The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Invitrogen Vector NTI Cloning & SequenceAnalysis Software (Ch. 4)BenchPro 2100 Plasmid Purification SystemInvitrogen protein expression vectorsincluding Gateway vectors (Ch. 6)E. coli, baculoviral, lentiviral, adenoviral, yeast,& mammalian expression systems (Ch. 6)Invitrogen Lipofectamine transfectionreagents (Ch. 6)Neon Transfection System (Ch. 6)MembranePro Functional Protein ExpressionKit (Ch. 6)Tali Image-Based Cytometer(information at www.lifetechnologies.com)Countess Automated Cell Counter(information at www.lifetechnologies.com)Invitrogen One Shot Competent Cells (Ch. 4)imMedia growth medium & LB broth (Ch. 4)Selection antibiotics (Ch. 6)

introductionintroductionYou may notice that we’ve included only our most popular products and featured new technologies within these pages.Follow the web link within the chapters for a complete view of all our products. While you’re on our website, be sure to checkout our new products, protocols, technical guides, videos, and handbooks at www.lifetechnologies.com.If you have any questions about which of our solutions is right for you, please contact Life Technologies technical support byphone, email, Facebook, or Twitter at www.facebook.com/LifeTechnologies and twitter.com/LifeTechSupport.Functional genetic analysisProtein analysis TRIzol Reagent (Ch. 2)Ambion Cells-to-Ct (Ch. 2)Ambion mirVana miRNA Isolation Kit (Ch. 2)Ambion mirVana PARIS Kit (Ch. 2)ArcturusXT Laser Capture Dissection System (Ch. 7) Dynabeads streptavidin magnetic beads(Ch. 6)Invitrogen Qubit Protein QuantitationSystem (Ch. 6)IP antibodies (Ch. 5) TaqMan and SYBR master mixes (Ch. 1)TaqMan Gene Expression, Genotyping (SNP), and CNVAssays (Ch. 1)TaqMan Non-coding RNA and Ambion Pre-miR Assays(Ch. 5)Epigenetics: methylation, histone/chromatin remodelingreagents (Ch. 5)Ambion Silencer Select, Lipofectamine RNAiMAXreagents, mirVana miRNA Mimics and Inhibitors (Ch. 5) Novex and NuPAGE Gels (Ch. 6)XCell SureLock gel box (Ch. 6)TaqMan Protein Assays (Ch. 1) iBlot Dry Blotting System(Ch. 6)BenchPro 4100 Western Processing Station(Ch. 6) Applied Biosystems Real-Time PCR Instruments StepOnePlus System & 7500 Fast System (Ch. 1)ViiA 7 Real-Time PCR System (Ch. 1)OpenArray System (Ch. 1) Applied Biosystems primers (Ch. 1)SYBR Green assays (Ch. 1)SuperScript VILO kits (Ch. 3) Novex Protein Stains (Ch. 6)Novex Buffers and protein standards (Ch. 6)Novex Gel cassettes andpour-your-own )Life Technologies offers products and services across many applications. Visit our website to find out more:Cell Culture solutions, go to www.lifetechnologies.com/cellculture.Cell and Tissue Analysis solutions, go to ng Solutions, go to logies.comIII

real-time PCR1

real-time PCR5Real-time PCR instruments8Real-time PCR instrument selection guide8Applied Biosystems OpenArray Real-Time PCR Platform911Applied Biosystems 7500 Fast Real-Time PCR System12Applied Biosystems 7500 Real-Time PCR System13StepOnePlus /StepOne Real-Time PCR Systems14Applied Biosystems 7900HT Fast Real-Time PCR System16High Resolution Melt (HRM) Software17RealTime StatMiner Software17Real-time PCR reagents and kits1chapter 1 contentsApplied Biosystems ViiA 7 Real-Time PCR Systemchapter sectionOverview of real-time PCR18Master mix selection guide18TaqMan Fast Advanced Master Mix19TaqMan Universal Master Mix II20TaqMan PreAmp Master Mix21TaqMan Sample-to-SNP Kit22TaqMan Genotyping Master Mix23MeltDoctor HRM Master Mix24Fast SYBR Green Master Mix25Power SYBR Green PCR Master Mix26Express One-Step SuperScript qRT-PCR Universal27TaqMan Fast Virus 1-Step Master Mix28Power SYBR Green RNA-to-Ct 1-Step Kit29TaqMan Exogenous Internal Positive Control Reagents30Other control reagents, enzymes, primers, and reagents31Spectral calibration kits33TaqMan RNase P Instrument Verification Plates34www.lifetechnologies.com2

real-time PCRchapter 1 contents13Gene expression35TaqMan Gene Expression Assay selection guide35TaqMan Gene Expression Assays36Lyophilized TaqMan Gene Expression Assays with Master Mix37Custom TaqMan Gene Expression Assays38Custom Plus TaqMan RNA Assays38TaqMan RNase P Detection Reagents39TaqMan Endogenous Controls39TaqMan Array Gene Signature Plates41Custom TaqMan Array Plates42TaqMan Custom Plating Service43Custom TaqMan Array Microfluidic Cards44TaqMan Array Gene Signature Microfluidic Cards45TaqMan OpenArray Real-Time PCR Plates46Custom TaqMan probes and primers48GeneAssist Pathway Atlas49Custom TaqMan Assay Design Tool49TaqMan MicroRNA and Non-coding RNA Assay selection guide50TaqMan MicroRNA Assays51Custom TaqMan Small RNA Assays51TaqMan Non-coding RNA Assays52TaqMan Pri-miRNA Assays52Megaplex Primer Pools53TaqMan MicroRNA Reverse Transcription Kit53TaqMan Array Human and Rodent MicroRNA Cards55TaqMan OpenArray MicroRNA Panels56TaqMan Protein Assays57DataAssist Software58ProteinAssist Software58

real-time PCRchapter sectionGenetic variation5959TaqMan SNP Genotyping Assays60Custom TaqMan SNP Genotyping Assays61TaqMan Drug Metabolism Genotyping Assays61GeneAssist Copy Number Assay Workflow Builder61TaqMan Copy Number Assays62TaqMan OpenArray Genotyping Plates64TaqMan OpenArray Digital PCR Kits66TaqMan Mutation Detection Assays68TaqMan Genotyper Software69CopyCaller Software69Custom TaqMan Assay Design Tool69www.lifetechnologies.com1chapter 1 contentsTaqMan SNP genotyping selection guide4

real-time PCROverview of real-time PCRAdvantages of real-time PCRQuantitative real-time PCR, also called qPCR, combinesPCR amplification and detection into a single step, thuseliminating the need to detect products using gel electrophoresis and enabling the PCR method to be truly quantitative. Real-time PCR systems utilize fluorescent dyes todetect the accumulation of PCR products during the exponential phase of the reaction, which allows for fast andprecise product quantification and objective data analysis.In contrast, end-point PCR requires subjective analysis ofproduct via gel electrophoresis, northern blots, or arrays.qPCR has several advantages over these semi-quantitativedetection methods, including:Real-time PCR1 Lower risk of contaminationEasier workflowFaster time-to-resultsBroad dynamic range: up to 9 logsHigher sensitivityEase of optimizationCost-effectivenessHigh throughputEasy-to-design controlsSmall input amounts of RNA or DNA requiredHigh reproducibilityTaqMan and SYBR Green detectionmethodsTaqMan probe–based detectionTaqMan chemistry entails the use of a fluorogenic probeto enable the detection of a specific PCR product as itRQ5'3'3'5'3'5'Step 1: Polymerization. A reporter (R) and a quencher (Q)are attached to the 5 and 3 ends of a TaqMan probe.FORWARDPRIMERRPROBEQForward and reverse primers are designed to hybridize tothe target sequence flanking either side of the probe. Atthe start of the PCR, in conjunction with the reverse primer,the probe attaches to the complementary region of thetarget DNA. As the enzyme begins extending the reverseprimer creating a new strand of DNA, it also degrades theattached probe that lies in its path. As the probe breaksapart, the reporter and quencher dye are separated. Theirloss of proximity, and thus quenching, results in a fluorescent signal from the reporter dye. The reporter dyesignal increases with each PCR cycle, and this signal isdirectly proportional to the amount of target DNA initiallypresent in the reaction. A single cycle of the TaqMan Assay is described in the figure below.SYBR Green–based detectionWhile SYBR Green–based detection offers less specificity than TaqMan probe–based approaches and doesnot enable assay multiplexing, it is often a less expensive method for qPCR analysis. However, more time istypically required to develop and optimize assays withQR5'5'3'accumulates during PCR. A single-stranded DNA probe isconstructed containing a reporter fluorescent dye on the5 end and a quencher dye on the 3 end. While the probeis intact, the proximity of the quencher dye greatly reducesthe fluorescence emitted by the reporter dye by fluorescence resonance energy transfer (FRET) through space. Ifthe target sequence is present, the probe anneals downstream from one of the primer sites and is cleaved by the5 nuclease activity of Taq DNA polymerase as this primeris extended.3'5'3'REVERSE PRIMER5'5'3'5'Step 2: Strand displacement. When both labels are attachedto the probe, reporter dye fluorescence is quenched.R REPORTERQ QUENCHER5'R5'5'3'3'5'5'Step 3: Cleavage. During each extension cycle, Taq DNApolymerase cleaves the reporter dye from the probe.3'Q3'5'3'5'Step 4: Polymerization completed. Once separated from thequencher, the reporter dye emits its characteristic fluorescence.TaqMan Assay. TaqMan chemistry uses a fluorogenic probe to enable the detection of a specific PCR product as it accumulates during PCR.5For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. 2011 Life Technologies Corporation. All rights reserved.The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.

real-time PCRStep 2: Denaturation. When the DNA is denatured, theSYBR Green I dye is released and fluorescence is drastically reduced.FORWARDPRIMERchapter sectionStep 1: Reaction setup. The SYBR Green I dye fluoresces when bound to double-stranded DNA.1Real-time PCRREVERSEPRIMERStep 3: Polymerization. During extension,primers anneal and PCR product is generated.Step 4: Polymerization completed. SYBR Green I dye bindsto the double-stranded product, resulting in a net increasein fluorescence detected by the instrument.SYBR Green Assay. SYBR Green I dye is nonspecific and will bind to any double-stranded DNA molecule.this approach. SYBR Green I is a dye that binds to theminor groove of double-stranded DNA. When bound, theintensity of its fluorescent emission increases. Hence, asmore double-stranded amplicons are produced duringPCR, the SYBR Green I dye signal increases proportionally (see figure above). The sensitivity of SYBR Green I exceeds that of ethidium bromide and otherconventional dyes by more than one order of magnitude.Researchers often use SYBR Green chemistry for initialscreening assays using qPCR. SYBR Green technologyis ideal for screening multiple targets or when a targetspecific probe is not readily available. These assays canlater be converted to TaqMan probe–based detectionsystems, which have much greater specificity. SYBR Green reagents include primers and master mixes thatcontain the SYBR Green I dye.Real-time PCR instrumentsReal-time PCR instruments are used to perform the PCRas well as the real-time detection and monitoring of theresulting fluorescent signal. The Applied Biosystems family of real-time PCR systems, which includes the ViiA 7, OpenArray , 7900HT Fast, 7500 Fast, 7500, StepOnePlus , and StepOne Real-Time PCR Systems, make qPCRmore accessible than ever. These systems offer cuttingedge software designed to support your application andare flexible, accommodating the real-time chemistry ofyour choice.Real-time PCR reagents and kitsLife Technologies’ optimized master mixes and kits providethe ideal solution for sensitive, reliable, and reproducibleresults. The master mixes are available in multiple formatsfor different applications. qPCR reagents include the samecomponents as traditional PCR, such as primers, enzymes,and dNTPs. However, TaqMan chemistry is distinguishedby the use of fluorescent dyes. TaqMan technology, basedon the 5 nuclease assay, offers the highest specificity andsensitivity. SYBR Green technology is ideal for screeningmultiple targets or when a target-specific probe is notreadily available.TaqMan Assays and custom probesLife Technologies offers predesigned, off-the-shelf, genespecific probe and primer sets and Custom TaqMan probes and primers manufactured to your desired targetsequences. All products use TaqMan probe–based chemistry and are designed for use on the suite of AppliedBiosystems real-time PCR systems. Together thesecomponents provide the gold standard in quantitative geneexpression and genotyping applications, offering greatsensitivity, specificity, reproducibility, and broad dynamicrange.www.lifetechnologies.com6

real-time PCRWeb resourcesReal-Time PCR (Applications and Technologies htmlReal-Time PCR Reagents Selection /REALTIMEPCR REAGENT PRODUCTReal-Time PCR Instruments Selection cr-instruments.html?ICID EDIOrd61Real-Time PCR vs. Traditional PCR vs. Digital vs-traditional-pcr.html?ICID EDI-Lrn2Real-Time PCR: Understanding Ct (Application -time PCRReal-Time PCR Learning -learn.htmlBibliography1.Bustin SA, Benes V, Garson JA et al. (2009) The MIQE Guidelines —Minimum Information for Publication of Quantitative Real-Time PCRExperiments. Clinical Chemistry 55(4):611-622.Real-time PCR technologyHiguchi R, Fockler C, Dollinger G, Watson R (1993) Kinetic PCR anal1.ysis: real-time monitoring of DNA amplification reactions. Biotechnology (NY) 11(9):1026-1030.2.Gibson UE, Heid CA, Williams PM (1996) A novel method for real-timequantitative RT-PCR. Genome Res 6(10):995-1001.3.Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real-time quantitative PCR. Genome Res 6(10):986-994.4.Wang T, Brown MJ (1999) mRNA quantification by real-time TaqMan polymerase chain reaction: validation and comparison with RNaseprotection. Anal Biochem 269(1):198-201.Kutyavin IV, Afonina IA, Mills A et al. (2000) 3 -minor groove binder5.DNA probes increase sequence specificity at PCR extension temperatures. Nucleic Acids Res 28(2):655-661.Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression6.data using real-time quantitative PCR and the 2(-Delta Delta CT)Method. Methods 25(4):402-408.High resolution meltingHerrman MG, Durtschi JD, Bromley KL et al. (2006) Amplicon DNA1.melting analysis for mutation scanning and genotyping: cross-platformcomparison of instruments and dyes. Clinical Chemistry 52(3): 494–503.Digital PCRVogelstein B, Kinzler KW (1999) Digital PCR. Proc Natl Acad Sci1.USA 96(16):9236–9241.Kalinina O, Brown J, Silver J (1997) Nanoliter scale PCR with TaqMan 2.detection. Nucleic Acids Res 25(10):1999–2004.MicroRNA analysis1.Chen C, Ridzon DA, Broomer AJ et al. (2005) Real-time quantificationof microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33(20):e179.2.Mestdagh P, Feys T, Bernard N et al. (2008) High-throughput stemloop RT-qPCR miRNA expression profiling using minute amounts ofinput RNA. Nucleic Acids Res 36(21):e143.3.Schmittgen TD, Lee EJ, Jiang J et al. (2008) Real-time PCR quantification of precursor and mature microRNA. Methods 44(1):31–38.4.Mestdagh P, Van Vlierberghe P, De Weer A et al. (2009) A noveland universal method for microRNA RT-qPCR data normalization.Genome Biol 10(6):R64.Allelic discrimination applications1.Campsall PA, Au NH, Prendiville JS (2004) Detection and geno-typingof varicella-zoster virus by TaqMan allelic discrimination real-timePCR. J Clin Microbiol 42(4):1409–1413.72.3.4.5.6.Chen X, Weaver J, Bove BA (2008) Allelic imbalance in BRCA1 andBRCA2 gene expression is associated with an increased breastcancer risk. Hum Mol Genet 17:1336–1348.Foster JT, Okinaka RT, Svensson R (2008) Real-time PCR assays ofsingle-nucleotide polymorphisms defining the major Brucella clades.J Clin Microbiol 46:296–301.Johnson VJ, Yucesoy B, Luster MI (2004) Genotyping of single nucleotide polymorphisms in cytokine genes using real-time PCR allelicdiscrimination technology. Cytokine 27(6):135 141.Tafe LJ, Belloni DR, Tsongalis GJ (2007) Detection of the C282Y and H63Dpolymorphisms associated with hereditary hemochromatosis using theABI 7500 fast real time PCR platform. Diagn Mol Pathol (2):112 115.Livak KJ (1999) Allelic discrimination using fluorogenic probes andthe 5 nuclease assay. Genet Anal 14(5-6):143–149.Clinical applicationsEspy MJ, Uhl JR, Sloan LM (2006) Real-time PCR in clinical microbiology:1.applications for routine laboratory testing. Clin Microbiol Rev 19(1):165 256.2.Ponchel F, Toomes C, Bransfield K et al. (2003) Real-time PCR basedon SYBR -Green I fluorescence: an alternative to the TaqMan assayfor a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. BMC Biotechnol 3:18.3.Skrzypski M (2008) Quantitative reverse transcriptase real-time polymerase chain reaction (qRT-PCR) in translational oncology: lungcancer perspective. Lung Cancer 59(2):147–54.4.Yoshida A, Kawada M, Suzuki N (2004) TaqMan real-time polymerase chain reaction assay for the correlation of Treponema denticola numbers with the severity of periodontal disease. Oral MicrobiolImmunol 19(3):196 200.5.He L, He X, Lim LP et al. (2007) A microRNA component of the p53tumour suppressor network. Nature 447(7148):1130–1134.6.Sharp AJ, Mefford HC, Li K et al. (2008) A recurrent 15q13.3 microdeletion syndrome associated with mental retardation and seizures.Nat Genet 40(3):322–328.7.Klein TE, Altman RB, Eriksson N et al. (2009) Estimation of the warfarindose with clinical and pharmacogenetic data. N Engl J Med 360(8):753–764.Forensic applications1.Evans JJ, Wictum EJ, Penedo MC et al. (2007) Real-time polymerasechain reaction quantification of canine DNA. Forensic Sci 52(1):93–96.2.Schulz MM, Schöbel K, Skowronek MH et al. (2005) Quantification ofhuman DNA from forensic samples via real time PCR by the help of atelomerase assay. Arch Kriminol 216(3-4):89 96.Swango KL, Timken MD, Chong MD et al. (2006) A quantitative PCR3.assay for the assessment of DNA degradation in forensic samples.Forensic Sci Int 158(1):14 26.4.Timken MD, Swango KL, Orrego C et al. (2005) A duplex real-timeqPCR assay for the quantification of human nuclear and mitochondrial DNA in forensic samples: implications for quantifying DNA indegraded samples. J Forensic Sci 50(5):1044 1066.Mutation allele detection1.Fujii K, Matsubara Y, Akanuma J et al. (2000) Mutation detection byTaqMan-allele specific amplification: application to molecular diagnosis of glycogen storage disease type Ia and medium-chain acylCoA dehydrogenase deficiency. Hum Mutat 15(2):189 196.Viral applications1.Fabre F, Kervarrec C, Mieuzet L et al. (2003) Improvement of barleyyellow dwarf virus-PAV detection in single aphids using a fluorescentreal time RT-PCR. J Virol Methods 110(1):51 60.2.He JW, Jiang S (2005) Quantification of enterococci and humanadenoviruses in environmental samples by real-time PCR. ApplEnviron Microbiol 71(5):2250 2255.3.Marroquí L, Estepa A, Perez L (2007) Assessment of the inhibitoryeffect of ribavirin on the rainbow trout rhabdovirus VHSV by real-timereverse-transcription PCR. Vet Microbiol 122(1-2):52 60.4.Rajendran KV, Cowley JA, McCulloch RJ et al. (2006) A TaqMan real-time RT-PCR for quantifying Mourilyan virus infection levels inpenaeid shrimp tissues. J Virol Methods 137(2):265 271.For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. 2011 Life Technologies Corporation. All rights reserved.The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.

real-time PCRReal-time PCR instruments selection guidechapter sectionApplied Biosystems real-time PCR instruments offer the gold standard for high-performance,multicolor real-time PCR. With these real-time PCR platforms, you get the convenience ofuniversal cycling conditions with TaqMan Assays and the flexibility of unlimited real-time chemistry choices.Instruments1StepOnePlus 96-well plates,0.1-mL tubesStepOne 48-well plates,0.1-mL tubesTemperatureuniformityFast block7500 Fast 96-welloptical Fast plates7500 96-well opticalplates or 0.2-mLtubes7900HT96-well, 384-well,and TaqMan ArrayMicrofluidic Cards 0.5 C, within30 sec of clockstart 0.5 C, within 30 secof clock start 0.5 C, within 30 secof clock startYesYes—7500 FastNo—7500Yes—96-well FastBlue LEDTungsten-halogen488 nm argon-ionlaserViiA 796-well, 384-well,and TaqMan ArrayMicrofluidic Cards 0.5 C, within30 sec of reachingtemperature setpointYes—96- and384-wellReal-time PCR instrumentsInstrument/capacityOpenArrat 3,072 through-holeplates 0.5 CNoOpticsExcitationsourceFilters/colorsStepOnePlus : 4filters factory-calibrated for FAM /SYBR Green,VIC /JOE , NED /TAMRA , andROX dyes6 excitation filters(450–670 nm)6 emission filters(500–720 nm)3 LEDs5 filter sets calibratedfor FAM /SYBR Green, VIC /JOE ,NED /TAMRA /Cy 3,ROX /Texas Red ,Cy 5 dyesNone, CCD acts asspectrograph withcontinuous detection500–650 nmCCDCCDCCDCCD10 copies10 copiesDown to 1 copy100 copiesUp to 9 logs of lineardynamic rangeUp to 9 logs of lineardynamic rangeUp to 9 logs of lineardynamic rangeUp to 6 logs of lineardynamic rangeW 23.6 cmD 51.2 cmH 42.7 cmW 34 cmD 45 cmH 49 cmw/ drawer open:W 72 cmD 84 cmH 64 cmW 53.3 cmD 63.5 cmH 64.5 cmW 63 cmD 77 cmH 72.5 cmYesYesStepOne : 3 filtersfactory-calibratedfor FAM /SYBR Green, VIC /JOE ,and ROX dyesDetectorOptiFlex Systemwith halogen lampPhotodiodesFilters are decoupled to allow for 21filter combinations3 filters factorycalibrated forFAM /SYBR Green, VIC /JOE ,and ROX dyesPerformance specificationsDemonstrated10 copiessensitivityDynamicUp to 9 logs of lineardynamic rangerangeInstrument .lifetechnologies.com8

real-time PCROpenArray Real-Time PCR PlatformGene expression, genotyping, microRNA, and digital PCR in a middensity, high-throughput formatReal-time PCR instruments1OpenArray Real-Time PCR Platform. Reliable, multiple sample loading—fill 3,072 through-holes (33 nL each) minimizing cross-contamination and withminimal hands-on time Economical, highly scalable approach—run up to three OpenArray Plates simultaneously on the OpenArray Real-TimePCR Instrument The OpenArray Real-Time PCR Platform provides PCR-based genotyping, gene expression, microRNA, and digital PCRanalysis of hundreds to thousands of samples and assays per day, enabling mid-density, high-throughput results withease of useComplete platformThe OpenArray Real-Time PCR Platform includes: OpenArray Real-Time PCR Instrument—images three OpenArray Plates simultaneouslyOpenArray AccuFill System—loads samples into OpenArray PlatesOpenArray Case Sealing Station—seals OpenArray PlatesOpenArray Instrument Accessories (includes PC system)OpenArray Real-Time PCR Analysis Software and OpenArray Genotyping Analysis SoftwareHundreds of samples and assaysResearchers can analyze up to 144 samples on a single OpenArray Plate. Three OpenArray Plates can be cycled andimaged simultaneously on the OpenArray Real-Time PCR Instrument, providing high-throughput results with ease. Thesystem workflow thermally cycles and fluorescently images the OpenArray Plates. Results are captured on the externalPC, and the OpenArray Analysis Software enables researchers to view specific assay data.9For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. 2011 Life Technologies Corporation. All rights reserved.The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.

real-time PCRWell-equipped cyclerAnalysis softwareOpenArray Platform technology is abroadly applicable nanoliter fluidicstechnology platform for low-volume,solution-phase reactions. Researchersusing this technology benefit from theparallelism of microarrays and the dataquality of solution-phase reactions suchas PCR.OpenArray Platform technology utilizesa microscope slide–sized plate with3,072 through-holes. Each through-holeis 300 µm in diameter with a depth of300 µm. The plates are arranged in 48subarrays of 64 through-holes. For geneexpression and genotyping applications,TaqMan or SYBR Assays (for up to 48samples) are preloaded and dried downin the plate, and are delivered ready forprocessing. For digital PCR applications,each through-hole is pre-treated toaccept your assays and samples in yourlab.OpenArray data can be easily manipulated and analyzed. Data frommutiple plates can be combined into the same project, and values suchas Ct, Tm, ΔCt, and ΔΔCt can be quickly calculated. Data can be analyzednumerically or graphically, and can be exported in .cvs format for furtheranalysis with third-party analysis packages.ProductOpenArray Real-Time PCR Instrument with QuantityCat. No.1 unit4459283AccuFill System OpenArray AccuFill System1 unit4457243OpenArray AccuFill System Tips1 box4457246OpenArray AccuFill System Tips, 10 pack10 boxes4458107OpenArray Loader Tips1 box4404571OpenArray Loader Tips, 10 pack10 boxes4404604OpenArray Case Sealing Station1 unit4409361Dual Flat Block GeneAmp PCR System 97001 unit4425757 Each OpenArray plate contains 48subarrays with 64 through-holes each.Proprietary processes are used to coatthe surfaces of the plates so that they arehydrophobic, while rendering the interiorsof the through-holes hydrophilic andbiocompatible. When processed, each ofthe 3,072 through-holes contains 33 nLof fluid held in place by means of surfacetension.Hydrophobic 1Real-time PCR instrumentsA number of software options make it easy to acquire, analyze, andmanage OpenArray data. OpenArray Real-Time PCR Software exportsgene expression data as Ct measurements. DataAssist Softwarepovides additional multi-plate functionality for advanced gene expression and miRNA analysis. For digital PCR, OpenArray Digital PCR Software fits real-time PCR data to a Poisson statistical model and displaysdigital results as copies per through-hole. OpenArray SNP GenotypingSoftware automates genotype calling, with easy export for further analysis. TaqMan Genotyper Software provides additional multi-plate functionality for powerful downstream analysis.OpenArray Platform Technologychapter sectionThe OpenArray Real-Time PCR Instrument uses three filtered highbrightness LEDs to excite, and a cooled CCD camera to detect fluorescence emission from dyes. The excitation and emission wavelengthssupport TaqMan FAM dye detection and SYBR Green I, ROX , FAM ,and VIC dye detection, making it an ideal platform for running allTaqMan and SYBR Green assays. The OpenArray Real-Time PCRInstrument includes an integrated thermal block

Molecular Biology Essentials Catalogue Comprising trusted and familiar Invitrogen and Applied Biosystems products, the Life Technologies molecular biology portfolio offers life sciences researchers the most-published, high-performance, scalable solutions across the breadth of molecular biology applications.

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