ORA™ qPCR Probe ROX H Mix
highQu qPCR mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qPCR cycling conditions. They provide excellent results on both AT and GC rich templates, in multiplexing and guaranty rapid extension with early Ct values with minimum or no optimization.
Detailní popis
Applications
- qPCR on instruments calibrated with high ROX concentration
- qPCR assays based on specific probes:
- TaqMan®, Molecular Beacons, Scorpions™ Probes
- Gene expression analysis, quantification of:
- gDNA, cDNA, viral DNA, low copy number genes
Benefits
- Universal - both standard and fast cycling, all probe qPCR assays
- qPCR of GC or AT rich templates, single-plex & multiplexing
- Rapid extension, early Ct
- Supplied with PCR Water
- Bulk quantities & custom packaging available
highQu qPCR mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qPCR cycling conditions. They provide excellent results on both AT and GC rich templates, in multiplexing and guaranty rapid extension with early Ct values with minimum or no optimization.
Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the ORA™ qPCR Probe Mastermixes are available in three versions – without ROX, with low or high ROX concentration.
For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qPCR Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software.
Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US.
Important Notes
- Use special primer selection programs for good planning.
- Work with amplicons in a range of 80-200, max 400 bp.
- Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.
- Run reactions in triplets; include a no-template control and positive control in parallel.
- Thaw and keep reagents on ice. Mix well before use.
Prepare a 20 µl reaction:
| Reverse Primer | 100 - 400 nM final c. |
| Forward Primer | 100 - 400 nM final c. |
| Specific Probe | 200 nM final c. |
| cDNA Template or gDNA Template |
<100 ng or <1 μg |
| PCR Water | to 10 μl |
| ORA™ qPCR Mix, 2X | 10 μl |
- Mix gently, avoid bubbles.
- Place into the instrument set like:
| Initial denaturation | 1 cycle: 95°C - 2 min for cDNA or 1 cycle: 95°C - 3 min for gDNA |
| Denaturation | 40 cycles: 95°C - 5 sec |
| Anneal./extension | 40 cycles: 60-65°C – 20-30 sec |
- Follow instrument instructions for melt curve analysis.
| CAT. | SIZE | COMPONENTS | COMPOSITION |
| QPP0301 | 200 r of 20 µl |
2 x 1 ml - ORA™ qPCR Probe ROX H Mix, 2X 2 x 1 ml - PCR Water |
Hot Start qPCR components: dNTPs at 0.25 mM, optimized buffer, low ROX concentration. |
| QPP0305 | 1000 r of 20 µl |
10 x 1 ml - ORA™ qPCR Probe ROX H Mix, 2X 10 x 1 ml - PCR Water |
Hot Start qPCR components: dNTPs at 0.25 mM, optimized buffer, low ROX concentration. |
Storage |
In the dark at -20°C. |
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