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Roche Applied ScienceFor general laboratory use. Not for use indiagnostic procedures. FOR IN VITRO USE ONLY.FastStart TaqMan ® Probe MasterFastStart TaqMan ®Probe Master (Rox)Cat. No. 04 673 409 001 2.5 ml (2 × 1.25 ml; 100 × 50 l reactions)Cat. No. 04 673 417 00112.5 ml (10 × 1.25 ml; 500 × 50 l reactions)Store at Ϫ15 to Ϫ25°CCat. No. 04 673 433 00150.0 ml (10 × 5 ml; 2000 × 50 l reactions)Cat. No. 04 673 450 001 2.5 ml (2 × 1.25 ml; 100 × 50 l reactions)Store at Ϫ15 to Ϫ25°C F Keep away from lightCat. No. 04 673 468 00112.5 ml (10 × 1.25 ml; 500 × 50 l reactions)Cat. No. 04 673 476 00150.0 ml (10 × 5 ml; 2000 × 50 l reactions)2× concentrated, ready-to-use hot start master mix for qPCR and qRT-PCR using the hydrolysis probe detection format on real-time PCR instruments (except the LightCycler ® Instruments)1.What this Product DoesContentsThe FastStart TaqMan ® Probe Master is a ready-to-use, 2× concen-trated master mix that contains all the reagents (except primers, probe and template) needed for running quantitative, real-time DNA detec-tion assays, including qPCR and 2-step qRT-PCR, in the hydrolysis probe detection format. It is available in two formulations, one that contains the Rox reference dye and one without Rox.Storage and StabilityIf stored at Ϫ15 to Ϫ25o C, the master mix is stable through the expira-tion date printed on the label. For short-term storage (up to 3months),the master mix may be stored at +2 to +8o C.N Keep the FastStart TaqMan ® Probe Master (Rox) away from light.N Avoid repeated freezing and thawing.L The complete PCR mix (i.e., FastStart TaqMan ® Probe Master sup-plemented with primers, probe, and template) is stable for up to 24hrs at room temperature. Keep the PCR mix away from light!ApplicationThe FastStart TaqMan ® Probe Master is a ready-to-use reagent mix that simplifies the preparation of reactions for qPCR and 2-step qRT-PCR. In combination with a real-time PCR instrument, suitable PCR primers and Hydrolysis Probe, FastStart TaqMan ® Probe Master allows very sensitive detection and quantification of defined DNA sequences.N Do not use this product on the LightCycler ® 1.5 Instrument, theLightCycler ® 2.0 Instrument, or the LightCycler ® 480 Instrument.In principle, the FastStart TaqMan ® Probe Master can be used for the amplification and detection of any DNA or cDNA target, including those that are GC-rich or GC-poor. However, you would need to adapt your detection protocol to the reaction conditions of the particular real-time PCR instrument in use and design a specific hydrolysis probe and PCR primers for each target. See the Operator’s Manual of your real-time PCR instrument for general recommendations.N The mix is designed for optimal amplification of targets up to500bp long. Do not use the mix to amplify longer targets.L FastStart TaqMan ® Probe Master offers convenience and ease-of-use because the addition of MgCl 2 to the reaction mixture is not necessary, thus avoiding time-consuming optimization steps.L The mix contains dUTP, so that it may be used with Uracil-DNAGlycosylase to prevent false positives arising from carry-over con-tamination, i.e., contamination with amplified DNA.L The FastStart TaqMan ® Probe Master is fully compatible withprobes from the Universal ProbeLibrary Sets*.Additional Equipment and Reagents RequiredAdditional reagents and equipment required to perform quantitative real-time PCR assays with the FastStart TaqMan ® Probe Master include:•general laboratory equipment–nuclease-free, aerosol-resistant pipette tips–pipettes with disposable, positive-displacement tips–sterile reaction tubes for preparing master mixes and dilu-tions–standard benchtop microcentrifuge –water, PCR-grade*•for first-strand cDNA synthesis–Transcriptor First Strand cDNA Synthesis Kit*•for real-time PCR–PCR reaction vessels (e.g., optical tubes or microplates)–sequence-specific primers–a hydrolysis probe (e.g ., from the Universal ProbeLibrary Sets*)–ROX Reference Dye* (optional)•for carry-over prevention (optional)–LightCycler ® Uracil-DNA Glycosylase** available from Roche Applied Science2.How to Use this Product2.1Before You BeginGeneralThe optimal reaction conditions (concentration of template DNA and PCR primers, incubation temperatures and times, cycle number)depend on the specific template/primer system and must be deter-mined individually.Sample Material•Use any template DNA (e.g., genomic or plasmid DNA, cDNA) suit-able for PCR in terms of purity, concentration, and absence of inhib-itors. For reproducible isolation of nucleic acids use:–either the MagNA Pure LC Instrument or the MagNA Pure Compact Instrument together with a dedicated nucleic acid isolation kit (for automated isolation) or–a HIGH PURE nucleic acid isolation kit (for manual isolation).0706.04699220001For details see the Roche Applied Science Biochemicals catalog or home page, .•Use up to 250 ng complex genomic DNA or 50 ng cDNA.PrimersUse PCR primers at a final concentration of 0.3 – 1.0 M. The recom-mended starting concentration is 0.9 M each.N Always use equimolar primer concentrations.N If you are using probes from the Universal ProbeLibrary*, use 200nM of each primer.L The design of the PCR primers determines amplicon length, melt-ing temperature, amplification efficiency, and yield. Primer design may also depend on the choice of PCR program (2-step versus 3-step protocol).L Several programs for primer design are freely available or provided by the suppliers of real-time PCR instruments (e.g., PrimerExpress).Alternatively, such programs are available to the public on the web for free. For example, use the free online ProbeFinder software () to design primers that may be paired with probes from the Universal ProbeLibrary*.ProbeThe probe concentration should be significantly lower than the primer concentration. As a starting point, we recommend using 250 nM probe. However, suitable concentrations range from 100 nM to 300nM.N To ensure a specific and sensitive assay, the probe must anneal to the DNA at a significantly higher temperature than the primers.Therefore, the T m of the probe should be 68 – 70°C and the T m of the primers about 58 – 60°C.N For maximum assay sensitivity, avoid placing a terminal G at the 5´-end of the probe because the fluorescent signal (arising after cleavage of the probe) is adversely affected by such a G.N To ensure that the fluorescent reporter dye within the unreacted probe is quenched, the length of the probe should not exceed 28 nucleotides.N If you use probes from the Universal Probe Library*, start with a probe concentration of 100 nM. Set the annealing temperature to 60°C.Negative ControlTo detect DNA contamination, always include a negative control in each run. To prepare this control, replace template DNA with PCR-grade water.Reaction VolumeVarious reaction volumes of the FastStart TaqMan® Probe Master can be used. Please refer to recommendations from the supplier of the instrument for suitable volumes and tubes/plates.ROX Reference DyeN Please read carefully.In principle, real-time PCR instruments (except the LightCycler®instruments) offer three different modes:1.Detection of released signal in relationship to a reference dye (usu-ally ROX)2.Detection of released signal in relationship to the quencher dye ofthe probe (e.g., TAMRA)3.Detection of released signal aloneThe choice of mode depends on the instrument (e.g. whether a chan-nel for detecting the reference dye is available) and on the light source of the instrument (halogen versus laser).The FastStart TaqMan® Probe Master is available with or without ROX. L The FastStart TaqMan® Probe Master (Rox) contains 120 nM ROX (2× concentrated), i.e. the final concentration of Rox in the PCR mix is 60 nM.The following list gives an overview how to apply the FastStart Taq-Man® Probe Master in combination with specific real-time PCR instru-ments:•If you want to use the reference channel of your real-time PCR instrument, then use the FastStart TaqMan® Probe Master (Rox).•for the ABI 7500 Real-Time PCR System, the Stratagene Mx3000P QPCR System, and the Corbett RotorGene Real Time Thermoana-lyzers: no addition of further ROX Reference Dye is required•for the ABI PRISM 7000 Sequence Detection System and the ABI7300 Real-Time PCR System: add additional ROX Reference Dyeto a final concentration of 300 nM a)•for the ABI 7700 Real-Time PCR System and the ABI PRISM7900HT Sequence Detection System: add additional ROX Refer-ence Dye to a final concentration of 400 nM a)•If you do not want to use the reference channel of your real-time PCR instrument or the instrument is not equipped with a reference channel, use either the FastStart TaqMan®Probe Master or the FastStart TaqMan® Probe Master (Rox).•If you use the Bio-Rad iCycler iQ5 Real-Time PCR Detection System apply only the External Well Factor Plate procedure for determining the well factors. For determining the well factors the Bio-Rad iCycler iQ5 Real-Time PCR Detection System does not use ROX but fluorescein. Therefore, use the FastStart TaqMan® Probe Mas-ter (without ROX) in combination with the iCycler iQ5 Real-Time PCR Detection System only! For details on how to perform the External Well Factor Plate procedure consult the iCycler iQ5 Real-Time PCR Detection System Instruction Manual.L a)This is the recommended starting concentration. The final con-centration of ROX Reference Dye may be further optimized in a range of 300 to 600 nM. Especially if you observe an unsteady flu-orescence signal, increase the ROX concentration until the signal is stable. Usually, when working with small reaction volumes (Յ10l) a higher ROX concentration is required. (In this case, the recommended starting concentration is 400 nM.)2.2ProcedurePreparation of the PCR mixFor each 50 l reaction, prepare the following reaction mix:ᕡ•Thaw the solutions and, for maximum recovery of the contents, briefly spin vials in a microcentrifuge before opening.•Mix solutions carefully by pipetting them up and down, thenstore on ice.ᕢPrepare 100× conc. solutions of the PCR primers and the hydrol-ysis probe.ᕣIn a 1.5 ml reaction tube on ice, prepare the PCR mix for one 50l reaction by adding the following components in the orderlisted below.Component Volume a)Finalconc.FastStart TaqMan® Probe MasterFastStart TaqMan® Probe Master (Rox)b25 l1×Hydrolysis Probe (25M)0.5 l250nMForward primer (90M)0.5 l900nMReverse primer (90M)0.5 l900nMWater, PCR-grade18.5 lT otal Volume45 lL a) To prepare the PCR mix for more than one reaction, multi-ply the amounts in the “Volume” column by z, where z = thenumber of reactions to be run + one additional reaction.N b) If you need to supplement the PCR mix with additional ROX Reference Dye*, add the dye to the FastStart TaqMan® ProbeMaster (Rox) before primers, probes or DNA template areadded. Refer to the package insert of the ROX Reference Dyefor detailed instructions.ᕤ•Mix the solution carefully by pipetting it up and down. Do not vortex.•Pipet45l PCR mix into each PCR reaction vessel or well of aPCR microplate (depending on your real-time PCR instrument).ᕥ•Add 5 l of template DNA (up to 250 ng total DNA) or cDNA.L In initial experiments to determine the optimum amount of cDNA template, we recommend running undiluted, 1:10diluted, and 1:100 diluted cDNA template in parallel.•Mix carefully by pipetting up and down.ᕦAccording to the instructions supplied with your instrument, prepare the tubes or microplates for PCR (e.g., seal tubes withoptical tube caps or the plate with self-adhesive foil).2Roche Applied Science3Roche Applied SciencePerforming PCRThere are several different ways to program the PCR. Either two-step or three-step PCR programs will provide suitable experimental results.The amplicon should be short (approx. 150 bp) and the annealing/elongation temperature should be 60°C (e.g., a typical PCR protocol is 40 cycles of 95°C/15 s, followed by 60°C/1 min).N For best results, be sure the instrument is calibrated correctly.2.3Related ProceduresPrevention of Carry-Over ContaminationUracil-DNA N-Glycosylase (UNG) is suitable for preventing carry-over contamination in PCR. This carry-over prevention technique involves incorporating deoxyuridine triphosphate into amplification products,then pretreating later PCR mixtures with UNG. If a dUTP-containing contaminant is present in the later PCRs, it will be cleaved by a combi-nation of the UNG and the high temperatures of the initial denatur-ation step; it will not serve as a PCR template. Since your target DNA template contains thymidine rather than uridine, it is not affected by this procedure.L dUTP is a component of the FastStart TaqMan ® Probe Master.N Perform prevention of carry-over contamination with LightCycler ®Uracil-DNA Glycosylase*. Add 1.25 U per 50 l PCR reaction. Pro-ceed as described in the package insert.Two-step RT-PCRFastStart TaqMan ® Probe Master can also be used to perform two-step RT-PCR. In two-step RT-PCR, the reverse transcription of RNA into cDNA is separated from the other reaction steps and is performed outside the real-time PCR instrument. Subsequent amplification and online monitoring is performed according to the standard real-time PCR procedure, using the cDNA as the starting sample material. Tran-scriptor First Strand cDNA Synthesis Kit* is recommended for reverse transcription of RNA into cDNA. Synthesis of cDNA is performed according to the detailed instructions provided with the kit.3.Troubleshooting³Following the instruction manual of the instrument supplier, program the instrument with the following parameters:CyclesAnalysis Mode Target Temperature Hold TimeRemarks 1 (optional)None 50°C2 minOnly if UNG hadbeen added for carry-over pre-vention 1None 95°C 10 minActivation of FastStart Taq DNA Polymerase 40Quantifi-cationDependent on the specific primer-probe combination.Amplification and real-time analysis·Place your tubes or plate in the instrument and start the reaction.»At the end of the reaction, follow instrument instructions for quantification/analysis.Problem CauseRecommendation No amplification detectable and no band in gel analysis•error in PCR program (e.g. activation step omitted)•pipetting errors (e.g. DNA not added)•amplicon too long •inhibitory effects of impurities•bad primer design•Adjust PCR program •Repeat experiment; check pipetting steps carefully•Redesign primer •Repeat isolation of template•Redesign primerNo or lowamplificationdetectable but strong band in gel analysisPCR is working but the probe is poorly designed Redesign probe 4.Additional Information on this ProductFastStart Taq DNA PolymeraseThe FastStart TaqMan ® Probe Master (with or without ROX) contains the FastStart Taq DNA Polymerase for hot-start PCR to improve speci-ficity and sensitivity of the PCR by minimizing the formation of non-specific amplification products (1,2,3,4). This enzyme delivers superior results thanks to its unique enzyme design and optimized buffer sys-tem.FastStart Taq DNA Polymerase is a chemically modified form of ther-mostable recombinant Taq DNA polymerase that shows no activity up to 75°C. The enzyme is active only at high temperatures, where primers no longer bind non-specifically. The enzyme is completely activated (by removal of blocking groups) in a single pre-incubation step (95°C,10 min) before cycling begins. Activation does not require the extra handling steps typical of other hot-start techniques.Detection of PCR productsReal-time DNA detection assays based on the hydrolysis probe format (also known as 5´-nuclease assays) require a single, signal-generating probe that contains two labels, a fluorescent reporter dye at the 5´-end and a (fluorescent or dark) quencher label at or near the 3´-end (6).When the probe is intact, the fluorescent signal is almost completely suppressed by the quenching label. When the probe is hybridized to its target sequence, it is cleaved by the 5´→3´ exonuclease activity of the FastStart Taq DNA Polymerase, which “unquenches” the fluores-cent reporter dye. During each PCR cycle, more of the released fluo-rescent dye accumulates, boosting the fluorescent signal.N If you use the hydrolysis probe format for detection, you cannotperform a subsequent melting curve analysis. For melting curve analysis, we recommend using the FastStart SYBR Green Master*.Quality ControlEach lot is tested for performance in qPCR using three templates:a GC–rich template, a GC-poor template and a long template (about 440 bp).References1Chou, Q et al (1992). Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nuc. Acid Res. 20, 1717-1723.2Kellogg, DE et al (1994). TaqStart Antibody: hot-start PCR facili-tated by a neutralizing monoclonal antibody directed against Taq DNA polymerase. BioTechniques 16, 1134-1137.3Birch, DE et al (1996). Simplified hot start PCR. Nature 381, 445-446.4PCR Manual, Roche Diagnostics (1999) 2nd edition (1999) 2, 52-58.5Bustin, SA (ed ., 2004). A – Z of Quantitative PCR. IUL Biotech-nology Series, 5.6Holland, PM et al (1991). Detection of specific polymerase chain reaction product by utilizing the 5´→3´ exonuclease activity of Thermus aquaticus DNA polymerase. Proc. Natl. Acad. Sci . USA . 88, 7276-7280.7Rees, E. et al (2006). siRNA Silencing of Lamin A and Quantifi-cation of the Knockdown Effect via qPCR. Biochemica 2006 (2), 25-27.Fluorescence varies within a run instrument not correctly calibratedRecalibrate instrument variations in pipettingMonitor the channel in which Rox is detected High background in the negative (no template) controlContamination •Remake or replace critical solutions (e.g ., water)•Clean lab bench •Use UNG to prevent carry-over contami-nationProblem Cause RecommendationRoche Diagnostics GmbHRoche Applied Science 68298 Mannheim Germany5.Supplementary Information5.1ConventionsText ConventionsTo make information consistent and memorable, the following text conventions are used in this package insert:SymbolsIn this package insert the following symbols are used to highlight important information:5.2AbbreviationsIn this Instruction Manual, the following abbreviations are used:5.3Changes to Previous Version•Storage and Stability information extended.•Recommended concentrations of ROX for ABI real-time PCR instruments changed.•Minor editorial changes5.4Ordering InformationRoche Applied Science offers a large selection of reagents and systems for life science research. For a complete overview of related products and manuals, please visit and book-mark our home page, , and our Special Interest Sites including:•The Universal ProbeLibrary: •The MagNA Pure System family for automated nucleic acid isolation: •Amplification – Innovative Tools for PCR: /pcr•DNA & RNA preparation – Versatile Tools for Nucleic Acid Purification: /napure L Refills for all 165 individual Universal ProbeLibrary probes, each sufficient for 500(50 l) reactions, can be ordered at .Text Convention UseNumbered Instructions labeled ቢ, ባ,etc.Steps in a process that usually occur in the order listed Numbered Instructions labeled ³, ·,etc.Steps in a procedure that must be performed in the order listed Asterisk *Denotes a product available from Roche Applied ScienceSymbolDescriptionL Information Note:Additional information about the current topic or procedure.NImportant Note:Information critical to the success of the procedure or use of the product.Abbreviation MeaningqPCR quantitative real-time PCR UNGUracil-DNA N-GlycosylaseProductPack SizeCat. No.FastStart SYBR Green Master (Rox) 5 ml (4 × 1.25 ml)50 ml (10 × 5 ml)04 673 514 00104 673 522 001FastStart SYBR Green Master 5 ml (4 × 1.25 ml)50 ml (10 × 5 ml)04 673 484 00104 673 492 001ROX Reference Dye50 l (1 mM)04 673 549 001Transcriptor First Strand cDNA Syn-thesis Kit1 kit 04 379 012 001LightCycler ® Uracil-DNA Glycosy-lase100 U 03 539 806 001Water, PCR Grade 25 ml (25 × 1 ml)25 ml (1 × 25 ml)100 ml (4 × 25 ml)03 315 932 00103 315 959 00103 315 843 001High Pure PCR T emplate Preparation Kit100 purifications 11 796 828 001mRNA Isolation Kit Ͼ70 g mRNA11 741 985 001Universal ProbeLibrary Set, Arabi-dopsis Library of 90 pre-validated detection probes04 683 595 001Universal ProbeLibrary Set, C. ele-gans Library of 90 pre-validated detection probes04 683 609 001Universal ProbeLibrary Set, Droso-phila Library of 90 pre-validated detection probes04 683 625 001Universal ProbeLibrary Set, Human Library of 90 pre-validateddetection probes04 683 633 001Universal ProbeLibrary Set, Mouse Library of 90 pre-validateddetection probes04 683 641 001Universal ProbeLibrary Set, Primates Library of 90 pre-validateddetection probes04 683 617 001Universal ProbeLibrary Set,Rat Library of 90 pre-validated detection probes04 683 650 001NOTICE TO PURCHASERLIMITED LICENSE: A license to perform the 5' nuclease process for research requires the use of a Licensed 5' Nuclease Kit (containing Licensed Probe), or the combination of an Authorized Core Kit plus Licensed Probe, or license rights that may be purchased from Applied Biosystems. This product is an Authorized Core Kit without Licensed Probe. Its purchase price includes a limited, non-transferable immu-nity from suit under U.S. Patents Nos. 5,210,015, 5,487,972, 5,476,774,and 5,219,727, and corresponding patent claims outside the United States, owned by Roche Molecular Systems, Inc. or F. Hoffmann-La Roche Ltd ("Roche"), for using only this amount of the product in the practice of the 5' nuclease process solely for the purchaser's own internal research and development activities. This product is also an Authorized Core Kit for use with service sublicenses available from Applied Biosystems. This product conveys no rights under U.S. Pat-ents Nos. 5,804,375, 6,214,979, 5,538,848, 5,723,591, 5,876,930,6,030,787, or 6,258,569, or corresponding patent claims outside the United States, expressly, by implication, or by estoppel.No right under any other patent claims (such as apparatus or system claims in U.S. Patent No. 6,814,934) and no right to perform commer-cial services of any kind, including without limitation reporting the results of purchaser's activities for a fee or other commercial consider-ation, is hereby granted expressly, by implication, or by estoppel. This product is for research purposes only. Diagnostic uses require a sepa-rate license from Roche.Further information on purchasing licenses to practice real-time PCR processes may be obtained by contacting the Director of Licensing,Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.TrademarksLIGHTCYCLER, FASTSTART, TAQMAN, HYBPROBE, MAGNA PURE,and HIGH PURE are Trademarks of Roche.ROX, PRISM, TAMRA, and PRIMER EXPRESS are Trademarks of Applera Corporation, Norwalk, CT, USA.Other brand or product names are trademarks of their respective holders.Contact and SupportTo ask questions, solve problems, suggest enhancements or report new applications, please visit our Online Technical Support Site at:/supportTo call, write, fax, or email us, visit the Roche Applied Science home page, , and select your home country. Country-specific contact information will be displayed.Use the Product Search function to find Pack Inserts and Material Safety Data Sheets.。
罗氏荧光定量PCR仪器简介罗氏荧光定量PCR仪器（Roche Real-Time PCR System）是一种用于荧光定量PCR（Polymerase Chain Reaction）的仪器。
PCR技术是一种广泛应用于分子生物学领域的技术，可以在体外大量复制DNA序列。
荧光定量PCR通过荧光信号的强度来定量PCR反应产物的数量，具有高灵敏度、高准确性和高通量的特点，被广泛应用于生命科学研究、疾病诊断和药物研发等领域。
罗氏是一家全球领先的医药和诊断解决方案提供商，旗下的荧光定量PCR仪器采用先进的技术和设计，为用户提供可靠和精确的PCR分析结果。
主要特点1.高灵敏度：罗氏荧光定量PCR仪器采用先进的光学系统，能够检测和定量非常低浓度的荧光信号，提高了实验的灵敏度。
2.高准确性：仪器内置的精确温度控制系统和优化的PCR反应条件，保证了实验结果的准确性和可重复性。
3.高通量：罗氏荧光定量PCR仪器支持多样品同时进行PCR反应，大大提高了实验的通量，节省了时间和成本。
4.灵活性：仪器配备了多样的PCR试剂和芯片，可以适应不同实验需求的扩增反应。
5.友好的用户界面：仪器具有直观的用户界面和易于操作的软件，用户可以轻松设置实验参数、监控实时反应和分析数据结果。
主要应用罗氏荧光定量PCR仪器在各种领域都有广泛的应用，包括但不限于以下几个方面：1.基因表达分析：通过定量PCR技术，可以准确测量基因在不同组织、细胞或疾病状态下的表达水平，帮助科研人员了解基因的功能和调控机制。
2.微生物检测：罗氏荧光定量PCR仪器可以用于检测和定量微生物如细菌、病毒、真菌等的核酸序列，广泛应用于食品安全、临床微生物学和环境监测等领域。
3.病毒载量检测：病毒载量是评估病毒感染严重程度的重要指标，罗氏荧光定量PCR仪器可以定量分析病毒的核酸含量，用于临床病毒学研究和病毒感染诊断。
4.药物研发：罗氏荧光定量PCR仪器可以用于药物研发过程中的基因表达分析、药物代谢相关基因的筛选和药物安全性评估等方面，为药物研发提供有力支持。
实时荧光定量PCR仪仪器原理及应用实时荧光定量PCR仪是一种基于PCR技术的分子生物学仪器。
它可以实时监测PCR 反应过程中的荧光信号，从而实现对靶标DNA的定量分析。
该仪器的原理基于原始的聚合酶链式反应（PCR）技术，通过PCR产生的DNA序列与特定荧光探针的结合，释放出荧光信号进行定量测量。
实时荧光定量PCR仪在生物医学研究、基因表达分析、病原体检测等方面具有广泛的应用。
通过该仪器，科研人员可以快速、准确地检测靶标DNA的含量，并获得相应数据用于后续分析。
仪器特点实时荧光定量PCR仪具有以下几个特点：1. 高灵敏度实时荧光定量PCR仪可以检测非常低浓度的靶标DNA，甚至达到单个分子的级别。
这一特点使得该仪器在医学诊断和疾病早期检测中具有很大潜力。
2. 高准确性该仪器采用荧光定量技术，能够实时监测PCR反应过程中的荧光信号，从而避免了传统PCR反应后的测量误差。
同时，实时荧光定量PCR仪还具有内标校正功能，能够准确地计算出靶标DNA的含量。
3. 广泛应用实时荧光定量PCR仪在生物医学领域有着广泛的应用。
它可以用于基因表达分析、病原体检测、遗传疾病的检测等方面。
同时，该仪器还可以应用于农业科学、环境监测、食品安全等领域。
使用方法使用实时荧光定量PCR仪进行实验需要以下步骤：1. 样本制备首先需要从待测样品中提取目标DNA，并进行纯化和扩增。
这一步骤可以根据具体实验目的选择不同的提取方法。
2. 反应液的配置将所需的PCR反应药物配置出合适的反应液。
其中包括靶标DNA的引物和荧光探针，以及聚合酶和其他辅助物质。
3. 仪器设置将PCR反应液加入到实时荧光定量PCR仪的孔板或管条中，然后将样品放入仪器中。
根据实验需要设置合适的温度和时间条件。
4. 实时检测启动仪器后，开始进行PCR反应和实时荧光检测。
仪器会不断地记录PCR反应过程中的荧光信号，并将数据输出保存。
5. 数据分析使用相应的分析软件对实时荧光定量PCR仪的输出数据进行处理和分析。
