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TAQuest™ qPCR Master Mix with Helixyte™ Green *Low 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 contains a low amount of 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 *Low 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 *Low 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 *Low 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 37 references: Citation Explorer
Development of cycling probe based real-time PCR methodology for influenza A viruses possessing the PA/I38T amino acid substitution associated with reduced baloxavir susceptibility.
Authors: Osada, Hidekazu and Chon, Irina and Phyu, Wint Wint and Wagatsuma, Keita and Nagata, Nobuo and Kawashima, Takashi and Sato, Isamu and Saito, Tadashi and Kodo, Naoki and Masaki, Hironori and Asoh, Norichika and Tsuchihashi, Yoshiko and Shirahige, Yutaka and Ono, Yasuhiko and Shimada, Yasushi and Hamabata, Hirotsune and Saito, Kousuke and Saito, Reiko
Journal: Antiviral research (2021): 105036
Authors: Osada, Hidekazu and Chon, Irina and Phyu, Wint Wint and Wagatsuma, Keita and Nagata, Nobuo and Kawashima, Takashi and Sato, Isamu and Saito, Tadashi and Kodo, Naoki and Masaki, Hironori and Asoh, Norichika and Tsuchihashi, Yoshiko and Shirahige, Yutaka and Ono, Yasuhiko and Shimada, Yasushi and Hamabata, Hirotsune and Saito, Kousuke and Saito, Reiko
Journal: Antiviral research (2021): 105036
Identification of a MarR subfamily that regulates arsenic resistance genes.
Authors: Yu, Yanshuang and Chen, Jichen and Li, Yuanping and Liang, Jinxuan and Xie, Zhenchen and Feng, Renwei and Alwathnani, Hend A and Rosen, Barry P and Grove, Anne and Chen, Jian and Rensing, Christopher
Journal: Applied and environmental microbiology (2021): AEM0158821
Authors: Yu, Yanshuang and Chen, Jichen and Li, Yuanping and Liang, Jinxuan and Xie, Zhenchen and Feng, Renwei and Alwathnani, Hend A and Rosen, Barry P and Grove, Anne and Chen, Jian and Rensing, Christopher
Journal: Applied and environmental microbiology (2021): AEM0158821
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
Method for detecting hepatitis B virus in blood plasma at low viral load using real-time PCR.
Authors: Serikova, E N and Semenov, A V and Ostankova, Yu V and Totolian, Areg A
Journal: Klinicheskaia laboratornaia diagnostika (2021): 59-64
Authors: Serikova, E N and Semenov, A V and Ostankova, Yu V and Totolian, Areg A
Journal: Klinicheskaia laboratornaia diagnostika (2021): 59-64
SARS-CoV-2 Detection using Real Time PCR by a Commercial Diagnostic Kit.
Authors: Roy, S and Paul, S K and Barman, T K and Ahmed, S and Haque, N and Mazid, R and Debnath, P and Roy, S A
Journal: Mymensingh medical journal : MMJ (2020): 596-600
Authors: Roy, S and Paul, S K and Barman, T K and Ahmed, S and Haque, N and Mazid, R and Debnath, P and Roy, S A
Journal: Mymensingh medical journal : MMJ (2020): 596-600
Page updated on November 21, 2024
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