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TAQuest™ FAST qPCR Master Mix with Helixyte™ Green *High ROX*
TAQuest™ FAST qPCR Master Mix with Helixyte™ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR. The master mix delivers results within 50 minutes for 40 cycles of PCR in a 20 uL reaction volume. The mix includes an optimized buffer containing dNTPs and our proprietary TAQuest™ FAST Hot Start Taq DNA Polymerase enzyme, an enzyme designed to allow instant hot start which minimizes non-specific product formation thus allowing room temperature reaction setup. Only template and target primers are required to run the desired PCR reactions. TAQuest™ FAST qPCR Master Mix with Helixyte™ Green 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 contains a high amount of ROX reference dye.
Example protocol
SAMPLE EXPERIMENTAL PROTOCOL
The following protocol can be used as a guideline.
Note Thaw the TAQuest™ FAST qPCR Master Mix with Helixyte™ Green *High ROX* at room temperature. Vortex qPCR Master Mix thoroughly before use.
Table 2.Thermal cycling parameters
Note Thaw the TAQuest™ FAST qPCR Master Mix with Helixyte™ Green *High 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™ FAST qPCR Master Mix with Helixyte™ Green *High 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 | 60 °C |
| Time (m:ss) | 0:10 | 0:20 | 0:30 |
References
View all 4 references: Citation Explorer
Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation.
Authors: Lista, Maria Jose and Matos, Pedro M and Maguire, Thomas J A and Poulton, Kate and Ortiz-Zapater, Elena and Page, Robert and Sertkaya, Helin and Ortega-Prieto, Ana M and Scourfield, Edward and O'Byrne, Aoife M and Bouton, Clement and Dickenson, Ruth E and Ficarelli, Mattia and Jimenez-Guardeño, Jose M and Howard, Mark and Betancor, Gilberto and Galao, Rui Pedro and Pickering, Suzanne and Signell, Adrian W and Wilson, Harry and Cliff, Penelope and Kia Ik, Mark Tan and Patel, Amita and MacMahon, Eithne and Cunningham, Emma and Doores, Katie and Agromayor, Monica and Martin-Serrano, Juan and Perucha, Esperanza and Mischo, Hannah E and Shankar-Hari, Manu and Batra, Rahul and Edgeworth, Jonathan and Zuckerman, Mark and Malim, Michael H and Neil, Stuart and Martinez-Nunez, Rocio Teresa
Journal: PloS one (2021): e0256813
Authors: Lista, Maria Jose and Matos, Pedro M and Maguire, Thomas J A and Poulton, Kate and Ortiz-Zapater, Elena and Page, Robert and Sertkaya, Helin and Ortega-Prieto, Ana M and Scourfield, Edward and O'Byrne, Aoife M and Bouton, Clement and Dickenson, Ruth E and Ficarelli, Mattia and Jimenez-Guardeño, Jose M and Howard, Mark and Betancor, Gilberto and Galao, Rui Pedro and Pickering, Suzanne and Signell, Adrian W and Wilson, Harry and Cliff, Penelope and Kia Ik, Mark Tan and Patel, Amita and MacMahon, Eithne and Cunningham, Emma and Doores, Katie and Agromayor, Monica and Martin-Serrano, Juan and Perucha, Esperanza and Mischo, Hannah E and Shankar-Hari, Manu and Batra, Rahul and Edgeworth, Jonathan and Zuckerman, Mark and Malim, Michael H and Neil, Stuart and Martinez-Nunez, Rocio Teresa
Journal: PloS one (2021): e0256813
Development of a multiplex TaqMan qPCR assay for simultaneous detection and differentiation of four DNA and RNA viruses from clinical samples of sheep and goats.
Authors: Xu, Xingang and Yang, Feng and Zhang, Qi and Xu, Ying and Huang, Jiali and Fu, Mingzhe and Zhang, Weimin
Journal: Journal of virological methods (2019): 58-64
Authors: Xu, Xingang and Yang, Feng and Zhang, Qi and Xu, Ying and Huang, Jiali and Fu, Mingzhe and Zhang, Weimin
Journal: Journal of virological methods (2019): 58-64
An improved RT-IPCR for detection of pyrene and related polycyclic aromatic hydrocarbons.
Authors: Meng, X Y and Li, Y S and Zhou, Y and Sun, Y and Qiao, B and Si, C C and Hu, P and Lu, S Y and Ren, H L and Liu, Z S and Qiu, H J and Liu, J Q
Journal: Biosensors & bioelectronics (2016): 194-199
Authors: Meng, X Y and Li, Y S and Zhou, Y and Sun, Y and Qiao, B and Si, C C and Hu, P and Lu, S Y and Ren, H L and Liu, Z S and Qiu, H J and Liu, J Q
Journal: Biosensors & bioelectronics (2016): 194-199
Evaluation and utilization of preassembled frozen commercial fast real-time qPCR master mixes for detection of cytomegalovirus and BK virus.
Authors: Glover, William A and Atienza, Ederlyn E and Nesbitt, Shannon and Kim, Woo J and Castor, Jared and Cook, Linda and Jerome, Keith R
Journal: Journal of medical virology (2016): 115-9
Authors: Glover, William A and Atienza, Ederlyn E and Nesbitt, Shannon and Kim, Woo J and Castor, Jared and Cook, Linda and Jerome, Keith R
Journal: Journal of medical virology (2016): 115-9
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