Real-Time PCR Enzymes & KitsProbe-Based One-Step qRT-PCR12Probe-Based qPCR13Reverse Transcription & cDNA Synthesis for qPCR8SYBR Green- or Dye-Based One-Step qRT-PCR6SYBR Green- or Dye-Based qPCR10QuantiTect Probe PCR KitsPrint For real-time PCR and two-step RT-PCR using sequence-specific probes Highly sensitive detection of low-copy targetsAccurate quantification over several logs of templateUse of any sequence-specific probe on any real-time cyclerAvailable with or without uracil-N-glycosylase (UNG)No need to optimize reaction and cycling conditions QuantiTect Probe PCR Kits enable sensitive quantification of gDNA and cDNA targets by real-time PCR and two-step RT-PCR using sequence-specific probes. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization. The dNTP mix includes dUTP, allowing optional treatment with UNG. For convenience, the master mix can be stored at 2–8°C. Buy ProductsProduct DetailsSpecificationsProduct ResourcesBuy ProductsCat No./ID:204343QuantiTect Probe PCR Kit (200)$479.00Order ProductFor 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 2 x 2 ml RNase-Free WaterCat No./ID:204345QuantiTect Probe PCR Kit (1000)$1,976.00Order ProductFor 1000 x 50 µl reactions: 25 ml 2x QuantiTect Probe PCR Master Mix, 20 ml RNase-Free WaterCat No./ID:204363QuantiTect Probe PCR +UNG Kit (200)$581.00Order ProductFor 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 100 ul UNG Solution, 2 x 2 ml RNase-Free WaterQuantiTect Probe PCR Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.See trademarks.Product Details Wide dynamic range in two-step PCR.Duplicate reactions were performed on the Mx3005P using tenfold dilutions of human keukocyte cDNA (10 ng to 0.01 ng) and a TaqMan assay for IL1R2 (a cytokine). The QuantiTect Probe PCR Kit provided accurate gene expression analysis from low to high template amounts with a PCR efficiency of 101%.High sensitivity and efficiency, and wide dynamic range.Tenfold serial dilutions of leukocyte cDNA (100 ng to 1 pg) were analyzed in duplicate on the Mx3005 using the QuantiTect Probe PCR Kit with primers and a FAM-labeled probe specific for IL8 (interleukin 8). A PCR efficiency of 97% over 6 logs of template dilution was achieved, and as little as 1 pg of IL8 transcript was sensitively detected.Specific primer annealing.Highly specific amplification.Wide dynamic range in real-time PCR.Probe-based real-time PCR with UNG pretreatment was carried out using the QuantiTect Probe PCR +UNG Kit. Reactions were run in duplicate on the ABI PRISM 7900 using 10-fold dilutions of human leukocyte cDNA (100 ng to 10 pg) and a TaqMan assay for IL8 (a cytokine). The amplification plots were evenly spaced, leading to a high PCR efficiency of 94%.Two-step RT-PCR.Effective UNG digestion.106 copies of two dUMP-containing PCR amplicons were treated with or without UNG and then amplified by real-time PCR. UNG was from various suppliers, and real-time PCR was performed using the master mix from the QuantiTect Probe PCR +UNG Kit. ΔCT on the Y-axis indicates CT values for non-UNG-treated samples subtracted from CT values for UNG-treated samples. The UNG supplied with the QuantiTect Kit provided the greatest ΔCT (9-12 cycles). This indicates that QIAGEN UNG digests carryover PCR products more effectively than UNG from other suppliers.Performance The QuantiTect Probe PCR Kit delivers sensitive and reliable results in combination with the QuantiTect Reverse Transcription Kit (see figure "High sensitivity and efficiency, and wide dynamic range"). The unique PCR buffer composition enables QuantiTect Probe PCR Kits to provide sensitive quantification of low-copy DNA targets, as well as accurate quantification over a wide linear range (see figures "High sensitivity and efficiency, and wide dynamic range" and "Wide dynamic range in two-step RT-PCR"). HotStarTaq DNA Polymerase further increases the specificity of the PCR reaction by providing the most stringent hot start compared with other polymerases (see figure "Highly specific amplification"). A major source of PCR contamination is the carryover of PCR products from previous reactions. Pretreatment with UNG prior to starting PCR ensures that any contaminating PCR products do not affect subsequent PCR. The combination of the specially optimized UNG solution and the proven PCR master mix in the QuantiTect Probe PCR +UNG Kit ensures effective elimination of carried-over PCR products together with reliable quantification of target sequences (see figures "Effective UNG digestion" and "Wide dynamic range in real-time PCR"). PrincipleQuantiTect Probe PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification of gDNA and cDNA targets using sequence-specific probes.The kits are designed for use with all types of sequence-specific probes, including hydrolysis probe detection (e.g., TaqMan® and other dual-labeled probes), FRET probes, and Molecular Beacons. QuantiTect Probe PCR Kits containa unique PCR buffer that containsa balanced combination of K+ and NH4+ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq DNA Polymerase provides a stringent hot start, preventingthe formation of nonspecific products. QuantiTect Probe PCR Master Mix also contains dUTP, enabling pretreatment with uracil-N-glycosylase (UNG) prior to starting PCR, which ensures that any contaminating PCR products do not affect subsequent PCR reactions. Components of 2x QuantiTect Probe PCR Kit Component Features Benefits HotStarTaq DNA Polymerase 15 minactivation at 95ºC Set-up of qPCR reactions at room temperature QuantiTect Probe PCR Buffer Balanced combination of NH4+ and K+ ions Specific primer annealing ensures reliable PCR results dNTP mix Includes dUTP, which partially replaces dTTP and enables optional UNG treatment of reactions Eliminatescontamination from carryover of PCR products by optional UNG treatment ROX dye For normalization of fluorescent signals on Applied Biosystems and, optionally, Agilent instruments Precise quantification on cyclers that require ROX dye.Does not interfere with reactions on other real-time cyclers Procedure QuantiTect Probe PCR Kits overcome the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers, probe, and DNA template to the ready-to-use PCR master mix, and start the reaction (see flowchart "Two-step RT-PCR"). Follow the protocol in the handbook to get fast and reliable results on any real-time cycler. If required, reactions can be pretreated with uracil-N-glycosylase (UNG) to eliminate carryover of PCR products from previous reactions.  For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiTect Reverse Transcription Kit. The kit provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination. Applications QuantiTect Probe PCR Kits can be used for probe-based gene expression analysis of cDNA targets or quantitative gDNA analysis on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent. For the Rotor-Gene Q and other Rotor-Gene cyclers, we recommend using the Rotor-Gene Probe PCR Kit, which has been specially developed for fast cycling on these instruments. SpecificationsFeaturesSpecifications ApplicationsReal-time quantification of DNA, cDNA, or RNA targets Reaction typePCR and two-step RT-PCR Real-time or endpointReal-time Sample/target typeDNA, cDNA, RNA Single or multiplexSingle SYBR Green I or sequence-specific probesSequence-specific probes Thermal cyclerMost real-time cyclers (e.g. LC, RG, ABI) With or without ROXWith ROXProduct ResourcesYou are not authorized to download the resourceBrochures & Guides (2) Sort optionsSort alphabetically (EN) - Analyzing Gene Expression and RegulationShow detailsCritical Factors for Successful Real-Time PCR - (EN)Show detailsShow allShow lessFAQs (30) Sort optionsSort alphabetically Do you recommend 1-step or 2-step real-time RT-PCR for gene expression analysis?FAQ ID -1056ViewWhat should I use as a standard for absolute quantification in real-time PCR?FAQ ID -1085ViewHow much template can I use in the reaction and what is the maximum volume of template that can be used in the QuantiTect Kits?FAQ ID -1086ViewHow much time will be saved when switching from standard cycling to fast cycling with QuantiFast Kits?FAQ ID -1438ViewHow do QuantiFast PCR Kits compare to QuantiTect PCR Kits for quantitative real-time PCR?FAQ ID -1441ViewWhy is the storage time for QuantiFast PCR Kits shorter than that for QuantiTect PCR Kits? FAQ ID -1446ViewWhy is the reaction volume for QuantiFast PCR Kits lower than that for QuantiTect PCR Kits?FAQ ID -1447ViewWhich real-time PCR kits are recommended downstream of the QuantiTect Whole Transcriptome Kit?FAQ ID -1592ViewDo the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment?FAQ ID -2117ViewWhat are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits?FAQ ID -2119ViewCan Uracil-N-Glycosylase (UNG) be purchased separately, independent of QuantiTect PCR Kits?FAQ ID -2134ViewAre QuantiTect PCR +UNG Kits available in bulk format (e.g., 25 ml master mix)?FAQ ID -2136ViewCan Uracil-N-Glycosylase (UNG) be added to the QuantiTect PCR Master Mix and be stored for later use?FAQ ID -2138ViewWill Uracil-N-Glycosylase (UNG) completely remove contaminating amplicons when using QuantiTect +UNG Kits?FAQ ID -2139ViewI would like to order the QuantiFast Probe Assays without master mix. Is this possible?FAQ ID -2360ViewDo you have any information or guidelines regarding the choice of reference genes for real-time PCR?FAQ ID -2371ViewWhat is the threshold cycle or Ct value? FAQ ID -2682ViewMy QuantiTect Master Mix did not freeze at degrees. Is it still ok to use? -20ViewWhy do replicates in real-time PCR have different plateau heights?FAQ ID -539ViewDo I need to calibrate my real-time cycler if I want to use Yakima Yellow?FAQ ID -540ViewWhat is a quenched FRET assay?FAQ ID -542ViewHow many template copies can I detect using QuantiTect Kits in real-time PCR?FAQ ID -543ViewWhat reaction volume should I use with QuantiTect Kits?FAQ ID -551ViewIs it possible to perform a UNG treatment when using QuantiTect kits?FAQ ID -564ViewCan I make a master mix with QuantiProbes, primers, and all other reaction components and store it for later use?FAQ ID -584ViewWhy are my realtime PCR amplification plots hook-shaped?FAQ ID -587ViewWhy does my realtime PCR assay quality decrease over time?FAQ ID -589ViewDo you have information on the use of recombinant DNA and RNA as absolute standards for realtime RT-PCR?FAQ ID -729ViewWhy should DNA or cDNA targets be less than 250 bp long for real-time PCR?FAQ ID-751ViewHow should fluorescent labeled probes be stored?FAQ ID -784ViewShow allShow lessKit Handbooks (2) Sort optionsSort alphabetically Uracil-N-glycosylase - (EN)For use with QuantiTect PCR Kits to eliminate carryover of PCR productsShow details(EN) - QuantiTect Probe PCR HandbookFor quantitative, real-time PCR and two-step RT-PCR using sequence-specific probesShow detailsShow allShow lessTechnical Information (1) Sort optionsSort alphabetically (EN) - Critical factors for synthesis of cDNA for real-time PCR analysisShow detailsShow allShow lessScientific Posters (1) Sort optionsSort alphabetically (EN) - Workflow for DNA purification from tough specimensShow detailsShow allShow lessQuick-Start Protocols (1) Sort optionsSort alphabetically QuantiTect Probe PCR Quick-Start ProtocolShow detailsShow allShow lessSafety Data Sheets (4) Sort optionsSort alphabetically MSDS QuantiTect Probe PCR Kit (200)MSDS QuantiTect Probe PCR Kit (1000)MSDS QuantiTect Probe PCR +UNG Kit (200)CofAShow allShow lessfragment fix placeholderProduct citations in scientific articlesPowered by Bioz See more details on BiozCustomers who bought these products also boughtCat No./ID:211392 Internal Control DNA (High conc.) For approximately 200 preps (depending on the elution volume): Lyophilized Internal Control DNA, Nucleic Acid Dilution Buffer$76.20Add To CartCat No./ID:622822QFT-Plus Ref Lab PackQFT-Plus Ref Lab PackAdd To CartCat No./ID:386113Matrix Standard BT5 single cap. (5 x 25)Matrix standard for 6-FAM, BTG, BTY, BTR, and BTO, for single-capillary analyzers$79.50Add To CartCat No./ID:211492 Internal Control RNA (High conc.) For approximately 200 preps (depending on elution volume): Lyophilized Internal Control RNA, Nucleic Acid Dilution Buffer$74.80Add To CartCat No./ID:204445QuantiTect Probe RT-PCR Kit (1000)For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Probe RT-PCR Master Mix, 0.5 ml QuantiTect RT Mix, 20 ml RNase-Free Water$2,763.00Add To CartCat No./ID:1117316DTT (1 ml)1M DTT, forensic grade quality; for sperm cell lysis.$90.00Add To CartCat No./ID:204443QuantiTect Probe RT-PCR Kit (200)For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe RT-PCR Master Mix, 100 µl QuantiTect RT Mix, 2 x 2 ml RNase-Free Water$672.00Add To CartCat No./ID:981301Rotor-Disc 72 (24)24 individually wrapped discs for 1728 reactions of 20–25 µl$264.00Add To CartCat No./ID:386123Matrix Standard BT5 multi cap. (25)Matrix standard for 6-FAM, BTG, BTY, BTR, and BTO, for multi-capillary analyzers$60.40Add To CartCat No./ID:211354QuantiFast Pathogen PCR +IC Kit (400)For 400 x 25 µl reactions: Master Mix, lyophilized Internal Control Assay, lyophilized Internal Control DNA, ROX Dye Solution, High-ROX Dye Solution, RNase-Free Water, Nucleic Acid Dilution Buffer, Buffer TE$945.00Add To CartCat No./ID:981601Rotor-Disc Heat Sealing Film (60)60 films for sealing Rotor-Disc 100 or Rotor-Disc 72 discs$120.00Add To CartCat No./ID:211454QuantiFast Pathogen RT-PCR +IC Kit (400)For 400 x 25 µl reactions: Master Mix, RT Mix, lyophilized Internal Control Assay, lyophilized Internal Control RNA, ROX Dye Solution, High-ROX Dye Solution, RNase-Free Water, Nucleic Acid Dilution Buffer, Buffer TE$1,129.00Add To CartCat No./ID:210212QIAGEN OneStep RT-PCR Kit (100)For 100 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 200 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 1 ml), dNTP Mix (1 x 200 µl, 10 mM each), 5x Q-Solution (1 x 2 ml), RNase-Free Water (2 x 1.9 ml)$615.00Add To CartCat No./ID:931955 QIAsymphony Certal Vaccine Kit (96) For 96 preps of 1000 µl each: includes 2 reagent cartridges and enzyme racks and accessories$1,508.00Add To CartCat No./ID:204574Rotor-Gene Probe RT-PCR Kit (400)For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Probe RT-PCR Master Mix, 100 µl Rotor-Gene RT Mix, 2 x 2 ml RNase-Free Water$689.00Add To CartCat No./ID:204843QuantiTect Multiplex RT-PCR NR Kit (200)For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Multiplex RT-PCR NoROX Master Mix (without ROX dye), 100 µl QuantiTect Multiplex RT Mix, 2 x 2 ml RNase-Free Water$1,006.00Add To CartCat No./ID:201912dNTP Set, PCR Grade, 4 x 100 µl100 mM each dATP, dCTP, dGTP, dTTP for 1000 x 50 µl PCR reactions$121.00Add To CartCat No./ID:929500QX Intensity Calibration Marker (1 ml)1 ml QX Intensity Calibration Marker$81.40Add To CartCat No./ID:1043369QIAamp DNA Blood Mini Accessory Set BAdditional Buffers and Reagents; Typically for use with 2x QIAamp DNA Blood Mini Kits (250), catalog number 51106, on the QIAcube$94.80Add To CartCat No./ID:204974Rotor-Gene Multiplex RT-PCR Kit (400)For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Multiplex RT-PCR Master Mix, 100 µl Rotor-Gene RT Mix, 2x2 ml RNase-Free Water$1,037.00Add To Cart