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TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX*

Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX*.
Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX*.
Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX*.
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Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


TAQuest™ qPCR Master Mix for TaqMan Probes is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR compatible with TaqMan gene expression assays. The master mix provides all of the essential components including our proprietary TAQuest™ Hot Start Taq DNA Polymerase enzyme and dNTPs in an optimized PCR buffer, except the template, primers and probes. The Hot Start Taq DNA polymerase allows you to set up a PCR reaction at room temperature, thus minimizing non-specific product formation. The optimized composition ensures PCR specificity and sensitivity with all sample types such as genomic, plasmid, viral and cDNA templates. TAQuest™ qPCR Master Mix for TaqMan Probes has been designed to be used for duplex reactions using internal positive controls. This master mix does not contain a ROX reference dye.



Instrument specification(s)Filter based on probes

Example protocol


The following protocol can be used as a guideline.
Note     Thaw the TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX* at room temperature. Vortex qPCR Master Mix thoroughly before use.
  1. Prepare one of the following reaction mixes as indicated in Table 1.
  2. Carefully mix the reagents with a gentle vortex followed by a brief centrifuge.
  3. Set up the plate in the qPCR instrument and run as indicated in Table 2. 
Table 1.Reagents composition per well for each reaction
Components Volume (25 µL/reaction) Volume (50 µL/reaction) Final Conc.
TAQuest™ qPCR Master Mix for TaqMan Probes *No ROX* 12.5 µL 25 µL 1X
Upstream primer, 10 µM 0.25-2.5 µL 0.5-5.0 µL 0.1-1.0 µM
Downstream primer, 10 µM 0.25-2.5 µL 0.5-5.0 µL 0.1-1.0 µM
TaqMan Probes, 10 µM 0.25-0.625 µL 0.5-1.25 µL 100-250 nM
DNA template 1-5 µL 1-5 µL Optimized conc.
Nuclease-Free Water to 25 µL 50 µL  
Table 2.Thermal cycling parameters
Parameter Polymerase Activation PCR (30-40 cycles)
  Hold Denature Anneal Extend
Temperature 95 °C 95 °C 55-65 °C 68-72 °C
Time (m:ss) 0:20 0:30 1:00 1:00



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Multiplex SYBR Green and duplex TaqMan real-time PCR assays for the detection of Photorhabdus Insect-Related (Pir) toxin genes pirA and pirB.
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sjTREC quantification using SYBR quantitative PCR for age estimation of bloodstains in a Japanese population.
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Quantification of M13 and T7 bacteriophages by TaqMan and SYBR green qPCR.
Authors: Peng, Xiujuan and Nguyen, Alex and Ghosh, Debadyuti
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Field Validation of SYBR Green- and TaqMan-Based Real-Time PCR Using Biopsy and Swab Samples To Diagnose American Tegumentary Leishmaniasis in an Area Where Leishmania (Viannia) braziliensis Is Endemic.
Authors: Gomes, Ciro Martins and Cesetti, Mariana Vicente and de Paula, Natália Aparecida and Vernal, Sebastián and Gupta, Gaurav and Sampaio, Raimunda Nonata Ribeiro and Roselino, Ana Maria
Journal: Journal of clinical microbiology (2017): 526-534
Qualitative Sybr Green real-time detection of single nucleotide polymorphisms responsible for target-site resistance in insect pests: the example of Myzus persicae and Musca domestica.
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Sybr Green- and TaqMan-Based Quantitative PCR Approaches Allow Assessment of the Abundance and Relative Distribution of Frankia Clusters in Soils.
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Authors: Dinoop, K P and Parija, Subhash Chandra and Mandal, Jharna and Swaminathan, R P and Narayanan, P
Journal: The Indian journal of medical research (2016): 49-56