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TAQuest™ qPCR Master Mix with Helixyte™ Green *High 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 high 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 *High 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 *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™ 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 | 55-65 °C | 68-72 °C |
| Time (m:ss) | 0:20 | 0:30 | 1:00 | 1:00 |
References
View all 7 references: Citation Explorer
SNPs and transcriptional activity of genes of innate and adaptive immunity at the maternal-fetal interface in woman with preterm labour, associated with preterm premature rupture of membranes.
Authors: Lyubomirskaya, Ekaterina S and Kamyshnyi, Alexandr M and Krut, Yuriy Ya and Smiianov, Vladyslav A and Fedoniuk, Larisa Ya and Romanyuk, Lidiya B and Kravets, Natalya Ya and Mochulska, Oksana M
Journal: Wiadomosci lekarskie (Warsaw, Poland : 1960) (2020): 25-30
Authors: Lyubomirskaya, Ekaterina S and Kamyshnyi, Alexandr M and Krut, Yuriy Ya and Smiianov, Vladyslav A and Fedoniuk, Larisa Ya and Romanyuk, Lidiya B and Kravets, Natalya Ya and Mochulska, Oksana M
Journal: Wiadomosci lekarskie (Warsaw, Poland : 1960) (2020): 25-30
Aligned Expression of IFI16 and STING Genes in RRMS Patients' Blood.
Authors: Helbi, Sobhan and Ravanbakhsh, Behnam and Karimi, Mohammad and Kooti, Wesam and Jivad, Nahid
Journal: Endocrine, metabolic & immune disorders drug targets (2020): 878-886
Authors: Helbi, Sobhan and Ravanbakhsh, Behnam and Karimi, Mohammad and Kooti, Wesam and Jivad, Nahid
Journal: Endocrine, metabolic & immune disorders drug targets (2020): 878-886
Real-Time Reverse Transcription PCR as a Tool to Study Virulence Gene Regulation in Bacterial Pathogens.
Authors: Aviv, Gili and Gal-Mor, Ohad
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 23-32
Authors: Aviv, Gili and Gal-Mor, Ohad
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 23-32
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
Analysis of P. gingivalis, T. forsythia and S. aureus levels in edentulous mouths prior to and 6 months after placement of one-piece zirconia and titanium implants.
Authors: Siddiqi, Allauddin and Milne, Trudy and Cullinan, Mary P and Seymour, Gregory J
Journal: Clinical oral implants research (2016): 288-94
Authors: Siddiqi, Allauddin and Milne, Trudy and Cullinan, Mary P and Seymour, Gregory J
Journal: Clinical oral implants research (2016): 288-94
Page updated on November 21, 2024
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