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TAQuest™ qPCR Master Mix with Helixyte™ Green *No ROX*
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.
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.
Table 2.Thermal cycling parameters
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.
| 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 |
| 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
A novel duplex SYBR Green real-time PCR with melting curve analysis method for beef adulteration detection.
Authors: Li, Jiapeng and Wei, Yixuan and Li, Jinchun and Liu, Ruixi and Xu, Suigen and Xiong, Suyue and Guo, Ya and Qiao, Xiaoling and Wang, Shouwei
Journal: Food chemistry (2021): 127932
Authors: Li, Jiapeng and Wei, Yixuan and Li, Jinchun and Liu, Ruixi and Xu, Suigen and Xiong, Suyue and Guo, Ya and Qiao, Xiaoling and Wang, Shouwei
Journal: Food chemistry (2021): 127932
Development and Validation of a SYBR Green Real Time PCR Protocol for Detection and Quantification of Nervous Necrosis Virus (NNV) Using Different Standards.
Authors: Olveira, José G and Souto, Sandra and Bandín, Isabel and Dopazo, Carlos P
Journal: Animals : an open access journal from MDPI (2021)
Authors: Olveira, José G and Souto, Sandra and Bandín, Isabel and Dopazo, Carlos P
Journal: Animals : an open access journal from MDPI (2021)
Simultaneous detection and differentiation of porcine circovirus 3 and 4 using a SYBR Green І-based duplex quantitative PCR assay.
Authors: Hou, Cheng-Yao and Xu, Tong and Zhang, Liu-Hui and Cui, Jian-Tao and Zhang, Yuan-Hang and Li, Xin-Sheng and Zheng, Lan-Lan and Chen, Hong-Ying
Journal: Journal of virological methods (2021): 114152
Authors: Hou, Cheng-Yao and Xu, Tong and Zhang, Liu-Hui and Cui, Jian-Tao and Zhang, Yuan-Hang and Li, Xin-Sheng and Zheng, Lan-Lan and Chen, Hong-Ying
Journal: Journal of virological methods (2021): 114152
Development of a SYBR Green-based RT-qPCR assay for the detection of Indian citrus ringspot virus.
Authors: Kokane, Amol D and Lawrence, Kapil and Kokane, Sunil B and Gubyad, Mrugendra G and Misra, Pragati and Reddy, M Krishna and Ghosh, Dilip Kumar
Journal: 3 Biotech (2021): 359
Authors: Kokane, Amol D and Lawrence, Kapil and Kokane, Sunil B and Gubyad, Mrugendra G and Misra, Pragati and Reddy, M Krishna and Ghosh, Dilip Kumar
Journal: 3 Biotech (2021): 359
A SYBR Green I-based real-time polymerase chain reaction assay for detection and quantification of canine bufavirus.
Authors: Wang, Yong and Sun, Jianfei and Guo, Xu and Li, Wei and Zhang, Da and Liu, Guangqing and Zhou, Tianhong and Li, Yongdong
Journal: Molecular and cellular probes (2021): 101762
Authors: Wang, Yong and Sun, Jianfei and Guo, Xu and Li, Wei and Zhang, Da and Liu, Guangqing and Zhou, Tianhong and Li, Yongdong
Journal: Molecular and cellular probes (2021): 101762
Page updated on November 20, 2024
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