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TAQuest™ FAST qPCR Master Mix with Helixyte™ Green *No 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 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™ FAST 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™ FAST 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™ FAST 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 | 60 °C |
| Time (m:ss) | 0:10 | 0:20 | 0:30 |
References
View all 10 references: Citation Explorer
Fatal systemic toxoplasmosis in a 3-month-old young tibetan goat (Capra hircus).
Authors: Pavone, Silvia and Crotti, Silvia and Cruciani, Deborah and D'Avino, Nicoletta and Zema, Jacopo and Morelli, Simone and Gobbi, Marco and Madeo, Laura
Journal: BMC veterinary research (2020): 423
Authors: Pavone, Silvia and Crotti, Silvia and Cruciani, Deborah and D'Avino, Nicoletta and Zema, Jacopo and Morelli, Simone and Gobbi, Marco and Madeo, Laura
Journal: BMC veterinary research (2020): 423
Development of four PCR-based methods to differentiate tilefish species (Branchiostegus japonicus and B. albus).
Authors: Kang, Tae Sun
Journal: Food chemistry (2019): 1-8
Authors: Kang, Tae Sun
Journal: Food chemistry (2019): 1-8
A rapid real-time PCR method to differentiate between mottled skate (Beringraja pulchra) and other skate and ray species.
Authors: Kim, Mi-Ra and Kwon, Kisung and Jung, Yoo-Kyung and Kang, Tae Sun
Journal: Food chemistry (2018): 112-119
Authors: Kim, Mi-Ra and Kwon, Kisung and Jung, Yoo-Kyung and Kang, Tae Sun
Journal: Food chemistry (2018): 112-119
A multi-screening Fast qPCR approach to the identification of abortive agents in ruminants.
Authors: Sebastiani, Carla and Curcio, Ludovica and Ciullo, Marcella and Cruciani, Deborah and Crotti, Silvia and Pesca, Cristina and Torricelli, Martina and Sebastianelli, Martina and Felici, Andrea and Biagetti, Massimo
Journal: Journal of microbiological methods (2018): 12-17
Authors: Sebastiani, Carla and Curcio, Ludovica and Ciullo, Marcella and Cruciani, Deborah and Crotti, Silvia and Pesca, Cristina and Torricelli, Martina and Sebastianelli, Martina and Felici, Andrea and Biagetti, Massimo
Journal: Journal of microbiological methods (2018): 12-17
Development of a Sensitive Real-Time Fast-qPCR Based on SYBR® Green for Detection and Quantification of Chicken Parvovirus (ChPV).
Authors: Nuñez, Luis F and Santander-Parra, Silvana H and Chaible, Lucas and De la Torre, David I and Buim, Marcos R and Murakami, Alexandre and Zaidan Dagli, Maria Lucia and Astolfi-Ferreira, Claudete S and Piantino Ferreira, Antonio J
Journal: Veterinary sciences (2018)
Authors: Nuñez, Luis F and Santander-Parra, Silvana H and Chaible, Lucas and De la Torre, David I and Buim, Marcos R and Murakami, Alexandre and Zaidan Dagli, Maria Lucia and Astolfi-Ferreira, Claudete S and Piantino Ferreira, Antonio J
Journal: Veterinary sciences (2018)
Page updated on November 21, 2024
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