TAQuest™ qPCR Master Mix for TaqMan Probes No ROX

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.

Example protocol

SAMPLE EXPERIMENTAL 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.

Prepare one of the following reaction mixes as indicated in Table 1.
Carefully mix the reagents with a gentle vortex followed by a brief centrifuge.
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

References

View all 50 references: Citation Explorer

Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples.
Authors: Pereira-Gómez, Marianoel and Fajardo, Álvaro and Echeverría, Natalia and López-Tort, Fernando and Perbolianachis, Paula and Costábile, Alicia and Aldunate, Fabián and Moreno, Pilar and Moratorio, Gonzalo
Journal: Journal of virological methods (2021): 114035

Lower cost alternatives for molecular diagnosis of COVID-19: conventional RT-PCR and SYBR Green-based RT-qPCR.
Authors: Dorlass, Erick Gustavo and Monteiro, Cairo Oliveira and Viana, Amanda Oliveira and Soares, Camila Pereira and Machado, Rafael Rahal Guaragna and Thomazelli, Luciano Matsumiya and Araujo, Danielle Bastos and Leal, Fabyano Bruno and Candido, Erika Donizette and Telezynski, Bruna Larotonda and Valério, Camila Araujo and Chalup, Vanessa Nascimento and Mello, Ralyria and Almeida, Flavia Jaqueline and Aguiar, Andressa Simões and Barrientos, Anna Carlotta Mott and Sucupira, Carolina and De Paulis, Milena and Sáfadi, Marco Aurélio Palazzi and Silva, Daniella Gregorio Bonfim Prado and Sodré, Janaina Joice Martins and Soledade, Mariana Pereira and Matos, Samantha Faria and Ferreira, Sabrina Rodrigues and Pinez, Célia Miranda Nunez and Buonafine, Carolina Palamin and Pieroni, Leticia Nery Ferreira and Malta, Fernanda Mello and Santana, Rubia Anita Ferraz and Souza, Eloisa Corrêa and Fock, Ricardo Ambrosio and Pinho, João Renato Rebelo and Ferreira, Luís Carlos Souza and Botosso, Viviane Fongaro and Durigon, Edison Luiz and Oliveira, Danielle Bruna Leal
Journal: Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] (2020): 1117-1123

Multiplex SYBR Green and duplex TaqMan real-time PCR assays for the detection of Photorhabdus Insect-Related (Pir) toxin genes pirA and pirB.
Authors: Cruz-Flores, Roberto and Mai, Hung Nam and Dhar, Arun K
Journal: Molecular and cellular probes (2019): 20-28

sjTREC quantification using SYBR quantitative PCR for age estimation of bloodstains in a Japanese population.
Authors: Yamanoi, Eisuke and Uchiyama, Saori and Sakurada, Makoto and Ueno, Yasuhiro
Journal: Legal medicine (Tokyo, Japan) (2018): 71-74

Quantification of M13 and T7 bacteriophages by TaqMan and SYBR green qPCR.
Authors: Peng, Xiujuan and Nguyen, Alex and Ghosh, Debadyuti
Journal: Journal of virological methods (2018): 100-107

Page updated on November 12, 2024

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Unit size	1 mL 5 mL
Catalog Number	1728217283
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Physical properties

Solvent

Water

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501