TAQuest™ qPCR Master Mix with Helixyte™ Green No ROX

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

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

Alternative formats

TAQuest™ qPCR Master Mix with Helixyte™ Green *Low ROX*

TAQuest™ qPCR Master Mix with Helixyte™ Green *High ROX*

Overview SDS Protocol

TAQuest™ qPCR Master Mix with Helixyte™ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR. The master mix includes our proprietary TAQuest™ Hot Start Taq DNA Polymerase enzyme and dNTPs in an optimized PCR buffer. You only need to add template and target primers to run the desired PCR reactions. The Hot Start Taq DNA polymerase allows you to set up a PCR reaction at room temperature, thus minimizing non-specific product formation. In combination with an optimized buffer, the enzyme ensures PCR specificity and sensitivity with all sample types such as genomic, plasmid, viral and cDNA templates. The Helixyte Green intercalating dye allows rapid DNA detection and analysis without using sequence-specific probes. This master mix does not contain a ROX reference dye.

Platform

qPCR

Instrument specification(s)

SYBR Green filter

Example protocol

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline.
Note Thaw the TAQuest™ qPCR Master Mix with Helixyte™ Green *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 with Helixyte™ Green 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
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

Images

Figure 1. Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix with Helixyte™ Green *No ROX*.

References

View all 50 references: Citation Explorer

Development and application of SYBR Green Ⅰ real-time quantitative reverse transcription PCR assay for detection of swine Getah virus.
Authors: Xia, Yin-He and Shi, Zi-Cong and Wang, Xin-Wei and Li, Yong-Tao and Wang, Zeng and Chang, Hong-Tao and Liu, Hong-Ying and Chen, Lu and Wang, Chuan-Qing and Yang, Xia
Journal: Molecular and cellular probes (2021): 101730

Development of SYBR Green I-based polymerase chain reaction for feline bocavirus 1 detection.
Authors: Wang, Yong and Li, Wei and Guo, Xu and Zhang, Da and Sun, Jianfei and Fu, Ziteng and Liu, Guangqing and Li, Yongdong and Jiang, Shudong
Journal: 3 Biotech (2021): 61

Duplex SYBR Green I-based real-time PCR assay for the rapid detection of canine kobuvirus and canine astrovirus.
Authors: Wang, Yong and Li, Yeqiu and Cui, Yongqiu and Jiang, Shudong and Liu, Hua and Wang, Jing and Li, Yongdong
Journal: Journal of virological methods (2021): 114066

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

Design and characterization of a SYBR Green I-based melting curve method for investigation of HER2I655V polymorphism in breast cancer.
Authors: Desriani, and Azamris, and Ghaissani, Shabrina S and Kinanti, Senja R and Warisman, Muhammad A and Fitria, N
Journal: Journal, genetic engineering & biotechnology (2021): 6

Development of SYBR Green I-based real-time reverse transcription polymerase chain reaction for the detection of feline astrovirus.
Authors: Wang, Yong and Fu, Ziteng and Guo, Xu and Zhang, Da and Bai, Caixia and Li, Wei and Liu, Guangqing and Li, Yongdong and Jiang, Shudong
Journal: Journal of virological methods (2021): 114012

A new SYBR Green real-time PCR to detect SARS-CoV-2.
Authors: Marinowic, D R and Zanirati, G and Rodrigues, F V F and Grahl, M V C and Alcará, A M and Machado, D C and Da Costa, J C
Journal: Scientific reports (2021): 2224

Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction.
Authors: Wang, Yong and Pan, Yang and Wu, Junhuang and Tong, Xinxin and Sun, Jianfei and Xu, Fazhi and Cheng, Bangzhao and Li, Yongdong
Journal: 3 Biotech (2021): 400

Establishment of Duplex SYBR Green I-based Real-time PCR Assay for Simultaneous Detection of Duck Hepatitis A Virus-1 and Duck Astrovirus-3.
Authors: Li, Yeqiu and Cui, Yongqiu and Liu, Hua and Wang, Jing and Li, Yongdong and Wang, Yong
Journal: Avian diseases (2021): 281-286

Validation of Swab Sampling and SYBR Green-Based Real-Time PCR for the Diagnosis of Cutaneous Leishmaniasis in French Guiana.
Authors: Blaizot, Romain and Simon, Stéphane and Ginouves, Marine and Prévot, Ghislaine and Blanchet, Denis and Ravel, Christophe and Couppie, Pierre and Demar, Magalie and Nabet, Cécile
Journal: Journal of clinical microbiology (2021)