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TAQuest™ FAST qPCR Master Mix for TaqMan Probes *Low 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 *Low 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 *Low 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 *Low ROX*.
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Physical properties
SolventWater
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
UNSPSC12171501

OverviewpdfSDSpdfProtocol


TAQuest™ FAST qPCR Master Mix for TaqMan Probes is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR well suited to use for TaqMan gene expression assays. The master mix is compatible with FAST conditions, thus delivers results within 50 minutes for 40 cycles of PCR in a 20 uL reaction volume. The master mix provides all of the essential components including our proprietary TAQuest™ FAST Hot Start Taq DNA Polymerase enzyme and dNTPs in an optimized PCR buffer, except the template, primers and probes. TAQuest™ FAST qPCR Master Mix for TaqMan Probes has been designed to be used for duplex reactions using internal positive controls with superior performance. The master mix ensures PCR specificity and sensitivity with all sample types such as genomic, plasmid, viral and cDNA templates. This master mix contains a low amount of ROX reference dye.

Platform


qPCR

Instrument specification(s)Filter based on probes

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline.
Note     Thaw the TAQuest™ FAST qPCR Master Mix for TaqMan Probes *Low 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
ComponentsVolume (25 µL/reaction)Volume (50 µL/reaction)Final Conc.
TAQuest™ FAST qPCR Master Mix for TaqMan Probes *Low ROX*12.5 µL25 µL1X
Upstream primer, 10 µM0.25-2.5 µL0.5-5.0 µL0.1-1.0 µM
Downstream primer, 10 µM0.25-2.5 µL0.5-5.0 µL0.1-1.0 µM
TaqMan Probes, 10 µM0.25-0.625 µL0.5-1.25 µL100-250 nM
DNA template1-5 µL1-5 µLOptimized conc.
Nuclease-Free Water to25 µL50 µL 
Table 2.Thermal cycling parameters
Parameter Polymerase Activation PCR (30-40 cycles)
  Hold Denature Anneal/Extend
Temperature 95 °C 95 °C 60 °C
Time (m:ss) 0:10 0:20 0:30

Images


References


View all 50 references: Citation Explorer
Evaluation of the efficiency of TaqMan duplex real-time PCR assay for non-invasive pre-natal assessment of foetal sex in equine.
Authors: Kadivar, Ali and Rashidzadeh, Habiballah and Davoodian, Najmeh and Nazari, Hasan and Dehghani Tafti, Rohallah and Heidari Khoei, Heidar and Seidi Samani, Hasan and Modaresi, Jahangir and Ahmadi, Ebrahim
Journal: Reproduction in domestic animals = Zuchthygiene (2021): 287-291
Fast and Sensitive Real-Time PCR Detection of Major Antiviral-Drug Resistance Mutations in Chronic Hepatitis B Patients by Use of a Predesigned Panel of Locked-Nucleic-Acid TaqMan Probes.
Authors: Chu, Son V and Vu, Son T and Nguyen, Hang M and Le, Ngan T and Truong, Phuong T and Vu, Van T T and Phung, Thuy T B and Nguyen, Anh T V
Journal: Journal of clinical microbiology (2021): e0093621
Multiplex TaqMan Real-Time PCR Assay for Sensitive Detection of Two Weevil Species (Coleoptera: Curculionidae).
Authors: Aguirre, Carlos and Sánchez, Evelyn and Olivares, Natalia and Hinrichsen, Patricio
Journal: Journal of economic entomology (2021): 90-99
A development strategy to fast establish the Taqman qPCR based method to detect SNP mutations.
Authors: Jiang, Xiaohui and Xiang, Junbei and Wang, Ruifeng and Wan, Qian
Journal: Human cell (2020): 1331-1333
One-step multiplex TaqMan probe-based method for real-time PCR detection of four canine diarrhea viruses.
Authors: Wang, Ruyi and Zhang, Wenyan and Ye, Rui and Pan, Zhongzhou and Li, Gairu and Su, Shuo
Journal: Molecular and cellular probes (2020): 101618
Development of a new TaqMan-based PCR assay for the specific detection and quantification of Simkania negevensis.
Authors: Baccari, Olfa and Elleuch, Jihen and Barkallah, Mohamed and Boukedi, Hanen and Ayed, Nourelhouda Ben and Hammami, Adnene and Fendri, Imen and Abdelkafi, Slim
Journal: Molecular and cellular probes (2020): 101645
Brief comparative evaluation of six open one-step RT-qPCR mastermixes for the detection of SARS-CoV-2 RNA using a Taqman probe.
Authors: Haddar, Cyrille and Verhoeven, Paul O and Bourlet, Thomas and Pozzetto, Bruno and Pillet, Sylvie
Journal: Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology (2020): 104636
BlueTYPE - A low density TaqMan-RT-qPCR array for the identification of all 24 classical Bluetongue virus serotypes.
Authors: Ries, Christina and Beer, Martin and Hoffmann, Bernd
Journal: Journal of virological methods (2020): 113881
Quantification of Major Bacteria and Yeast Species in Kefir Consortia by Multiplex TaqMan qPCR.
Authors: Nejati, Fatemeh and Junne, Stefan and Kurreck, Jens and Neubauer, Peter
Journal: Frontiers in microbiology (2020): 1291
Rapid identification of Babesia canis and Babesia gibsoni (Asian genotype) in canine blood samples using a customized portable real-time PCR analyzer and TaqMan-based assay.
Authors: Kuo, Chun-Yen and Zhao, Chihyu and Cheng, TsunLi and Tsou, Chih-Cheng and Li, Yi-Chen and Zhang, Yong and Hsieh, Ming-Che and Haung, Song-Bin and Chen, Wen-Ying
Journal: Ticks and tick-borne diseases (2020): 101362